Quaternary salt CCR2 antagonists

ABSTRACT

Quaternary salt compounds of Formula (I) 
                         
or pharmaceutically acceptable forms thereof, which are CCR2 antagonists and are useful in preventing, treating or ameliorating CCR2 mediated inflammatory syndromes, disorders or diseases in a subject in need thereof.

This application claims the benefit of provisional application Ser. No.60/582,929 filed on Jun. 24, 2004.

BACKGROUND OF THE INVENTION

The invention is directed to quaternary salt compounds which areantagonists to the chemoattractant cytokine receptor 2 (CCR2),pharmaceutical compositions, and methods for use thereof. Moreparticularly, the CCR2 antagonists are phenylamino substitutedquaternary salt compounds used in ameliorating or treating CCR2 mediatedinflammatory disorders.

CCR2 is a member of the GPCR family of receptors, as are all knownchemokine receptors and are expressed by monocytes and memoryT-lymphocytes. The CCR2 signaling cascade involves activation ofphospholipases (PLCβ₂), protein kinases (PKC), and lipid kinases (PI-3kinase).

Chemoattractant cytokines (i.e., chemokines) are relatively smallproteins (8-10 kD) which stimulate the migration of cells. The chemokinefamily is divided into four subfamilies based on the number of aminoacid residues between the first and second highly-conserved cysteines.

Monocyte chemotactic protein-1 (MCP-1) is a member of the CC chemokinesubfamily (wherein CC represents the subfamily having adjacent first andsecond cysteines) and binds to the cell-surface chemokine receptor 2(CCR2). MCP-1 is a potent chemotactic factor which, after binding toCCR2, mediates monocyte and lymphocyte migration (i.e., chemotaxis)toward a site of inflammation. MCP-1 is also expressed by cardiac musclecells, blood vessel endothelial cells, fibroblasts, chondrocytes, smoothmuscle cells, mesangial cells, alveolar cells, T-lymphocytes,marcophages, and the like.

After monocytes enter the inflammatory tissue and differentiate intomacrophages, monocyte differentiation provides a secondary source ofseveral proinflammatory modulators, including tumor necrosis factor-α(TNF-α), interleukin-1 (IL-1), IL-8 (a member of the CXC chemokinesubfamily, wherein CXC represents one amino acid residue between thefirst and second cysteines), IL-12, arachidonic acid metabolites (e.g.,PGE₂ and LTB₄), oxygen-derived free radicals, matrix metalloproteinasesand complement components.

Animal model studies of chronic inflammatory diseases have demonstratedthat inhibition of binding between MCP-1 and CCR2 by an antagonistsuppresses the inflammatory response. The interaction between MCP-1 andCCR2 has been implicated (see Rollins B J, Monocyte chemoattractantprotein 1: a potential regulator of monocyte recruitment in inflammatorydisease, Mol. Med. Today, 1996, 2:198; and Dawson J, et al., Targetingmonocyte chemoattractant protein-1 signaling in disease, Expert Opin.Ther. Targets, 2003 February, 7(1):35-48) in inflammatory diseasepathologies such as uveitis, atherosclerosis, rheumatoid arthritis,multiple sclerosis, Crohn's Disease, nephritis, organ allograftrejection, fibroid lung, renal insufficiency, diabetes and diabeticcomplications, diabetic nephropathy, diabetic retinopathy, diabeticretinitis, diabetic microangiopathy, tuberculosis, sarcoidosis, invasivestaphylococcia, inflammation after cataract surgery, allergic rhinitis,allergic conjunctivitis, chronic urticaria, allergic asthma, periodontaldiseases, periodonitis, gingivitis, gum disease, diastoliccardiomyopathies, cardiac infarction, myocarditis, chronic heartfailure, angiostenosis, restenosis, reperfusion disorders,glomerulonephritis, solid tumors and cancers, chronic lymphocyticleukemia, chronic myelocytic leukemia, multiple myeloma, malignantmyeloma, Hodgkin's disease, and carcinomas of the bladder, breast,cervix, colon, lung, prostate, or stomach.

Monocyte migration is inhibited by MCP-1 antagonists (either antibodiesor soluble, inactive fragments of MCP-1) which have been shown toinhibit the development of arthritis, asthma, and uveitis. Both MCP-1and CCR2 knockout (KO) mice have demonstrated that monocyte infiltrationinto inflammatory lesions is significantly decreased. In addition, suchKO mice are resistant to the development of experimental allergicencephalomyelitis (EAE, a model of human MS), cockroach allergen-inducedasthma, atherosclerosis, and uveitis. Rheumatoid arthritis and Crohn'sDisease patients have improved during treatment with TNF-α antagonists(e.g., monoclonal antibodies and soluble receptors) at dose levelscorrelated with decreases in MCP-1 expression and the number ofinfiltrating macrophages.

MCP-1 has been implicated in the pathogenesis of seasonal and chronicallergic rhinitis, having been found in the nasal mucosa of mostpatients with dust mite allergies. MCP-1 has also been found to inducehistamine release from basophils in vitro. During allergic conditions,both allergens and histamines have been shown to trigger (i.e., toup-regulate) the expression of MCP-1 and other chemokines in the nasalmucosa of people with allergic rhinitis, suggesting the presence of apositive feedback loop in such patients.

There remains a need for small molecule CCR2 antagonists for preventing,treating or ameliorating a CCR2 mediated inflammatory syndrome, disorderor disease resulting from MCP-1 induced monocyte and lymphocytemigration to a site of inflammation.

All documents cited herein are incorporated by reference.

SUMMARY OF THE INVENTION

The invention provides quaternary salt compounds of Formula (I)

or pharmaceutically acceptable forms thereof, which are CCR2 antagonistsand are useful in preventing, treating or ameliorating CCR2 mediatedinflammatory syndromes, disorders or diseases in a subject in needthereof.

The present invention also provides a method for preventing, treating orameliorating a CCR2 mediated inflammatory syndrome, disorder or diseasein a subject in need thereof comprising administering to the subject aneffective amount of a compound of Formula (I) or composition ormedicament thereof.

DETAILED DESCRIPTION OF THE INVENTION

The present invention is directed to a compound of Formula (I)

and pharmaceutically acceptable forms thereof, wherein

-   A is carbonyl, thiocarbonyl or sulfonyl;-   X is a bond or —CH═CH—;-   R₁ is selected from-   (1). aryl optionally substituted by one or more lower alkyl,    —(CH₂)_(n)—CF₃, lower alkoxy, alkoxycarbonyl, cyano, halogen or    phenyl optionally substituted by lower alkyl, —(CH₂)_(n)—CF₃, lower    alkoxy, alkoxycarbonyl, cyano or halogen;-   (2). C₅-C₁₅ cycloalkyl optionally substituted by one or more lower    alkyl, —(CH₂)_(n)—CF₃, lower alkoxy, aryl, halogen-substituted aryl,    alkoxycarbonyl, cyano or halogen; or,-   (3). heterocyclyl optionally substituted by one or more lower alkyl,    —(CH₂)_(n)—CF₃, lower alkoxy, aryl, aryl-lower alkyl,    halogen-substituted aryl, alkoxycarbonyl, cyano or halogen;-   n is 0, 1, 2, 3 or 4;-   Y is a bond or —CH₂—;-   X₂ is —(CH₂)_(m)— wherein m is 1 or 2;-   R₂ is —N⁺(R₄R₅)—ZR₃;-   Z is —(CH₂)_(p)— wherein p is 0, 1 or 2;-   R₃ is selected from-   (1). aryl optionally substituted with one or more lower alkyl,    —(CH₂)_(n)—CF₃, lower alkoxy, aryl, halogen-substituted aryl,    alkoxycarbonyl, cyano or halogen;-   (2). C₅-C₁₅ cycloalkyl optionally substituted with one or more lower    alkyl, —(CH₂)_(n)—CF₃, lower alkoxy, aryl, halogen-substituted aryl,    alkoxycarbonyl, cyano or halogen; or,-   (3). heterocyclyl optionally substituted with one or more lower    alkyl, —(CH₂)_(n)—CF₃, lower alkoxy, aryl, halogen-substituted aryl,    alkoxycarbonyl, cyano or halogen; wherein, when heterocyclyl is    attached via a carbon atom ring member and a heteroatom ring member    is adjacent to said carbon atom, then p is 1 or 2;-   R₄ and R₅ are each individually lower alkyl or lower alkenyl;-   alternatively, R₄ and R₅ combine with the nitrogen atom of    Formula (I) to form a heterocyclyl ring of 5 to 9 total ring atoms    optionally containing one of an oxygen or sulfur ring atom, wherein    the heterocyclyl ring nitrogen atom is substituted with one of lower    alkyl or lower alkenyl to form a quaternary salt, and wherein —ZR₃    is absent and the heterocyclyl ring is optionally substituted with    aryl optionally substituted with one or more lower alkyl,    —(CH₂)_(n)—CF₃, lower alkoxy, aryl, halogen-substituted aryl,    alkoxycarbonyl, cyano or halogen.

An example of the invention is a compound of Formula (I) andpharmaceutically acceptable forms thereof, wherein A is carbonyl; X is abond; R₁ is selected from aryl substituted by one or more lower alkyl orhalogen, C₅-C₁₅ cycloalkyl optionally substituted by one or morehalogen, or heterocyclyl optionally substituted by one or more loweralkyl or halogen; Y is a bond; X₂ is —CH₂—; R₂ is —N⁺(R₄R₅)—R₃; R₃ isselected from C₅-C₁₅ cycloalkyl or heterocyclyl and R₄ and R₅ are eachindividually lower alkyl.

An example of the invention is a compound of Formula (I) andpharmaceutically acceptable forms thereof, wherein A is carbonyl, X is abond, R₁ is aryl optionally substituted by one or more halogen, Y is abond, X₂ is —CH₂—, R₂ is —N⁺(R₄R₅)—R₃, R₃ is heterocyclyl and R₄ and R₅are each individually lower alkyl.

An example of the invention is a compound of Formula (I) andpharmaceutically acceptable forms thereof, wherein A is carbonyl.

An example of the invention is a compound of Formula (I) andpharmaceutically acceptable forms thereof, wherein R₁ is selected from

-   (1). aryl optionally substituted by one or more lower-alkyl,    —(CH₂)_(n)—CF₃, lower alkoxy, cyano, halogen or phenyl optionally    substituted by lower alkyl, —(CH₂)_(n)—CF₃, lower alkoxy, cyano or    halogen;-   (2). C₅-C₁₅ cycloalkyl optionally substituted by one or more lower    alkyl, —(CH₂)_(n)—CF₃, lower alkoxy, cyano or halogen; or,-   (3). heterocyclyl optionally substituted by one or more lower alkyl,    —(CH₂)_(n)—CF₃, lower alkoxy, aryl, aryl-lower alkyl,    halogen-substituted aryl or halogen.

An example of the invention is a compound of Formula (I) andpharmaceutically acceptable forms thereof, wherein n is 0.

An example of the invention is a compound of Formula (I) andpharmaceutically acceptable forms thereof, wherein p is 0 or 1.

An example of the invention is a compound of Formula (I) andpharmaceutically acceptable forms thereof, wherein R₃ is C₅-C₁₅cycloalkyl or heterocyclyl; wherein, when heterocyclyl is attached via acarbon atom ring member and a heteroatom ring member is adjacent to saidcarbon atom, then p is 1.

An example of the invention is a compound of Formula (I) andpharmaceutically acceptable forms thereof, wherein R₄ and R₅ are eachindividually lower alkyl or lower allyl.

An example of the invention is a compound of Formula (I) andpharmaceutically acceptable forms thereof, wherein R₄ and R₅ combinewith the nitrogen atom of Formula (I) to form a heterocyclyl ring of 5to 9 total ring atoms optionally containing one of an oxygen or sulfurring atom, wherein the heterocyclyl ring nitrogen atom is substitutedwith lower alkyl to form a quaternary salt, and wherein —ZR₃ is absentand the heterocyclyl ring is optionally substituted with aryl optionallysubstituted with one or more lower alkyl, —(CH₂)_(n)—CF₃, lower alkoxy,cyano or halogen.

An example of the invention is a compound of Formula (I) andpharmaceutically acceptable forms thereof, wherein R₄ and R₅ combinewith the nitrogen atom of Formula (I) to form a heterocyclyl ring of 5to 9 total ring atoms optionally containing one of an oxygen or sulfurring atom, wherein the heterocyclyl ring nitrogen atom is substitutedwith lower alkyl to form a quaternary salt, and wherein —ZR₃ is absentand the heterocyclyl ring is optionally substituted with aryl optionallysubstituted with lower alkoxy.

An example of the invention is a compound of Formula (Ia)

or pharmaceutically acceptable form thereof, wherein R₁, X, Y and X₂R₂are dependently selected from

Cpd R₁ X Y X₂R₂ 1 3-Br-phenyl —CH═CH— —CH₂— 4-CH₂—N⁺(CH₃)₂-cyclohexyl, 23-Br-phenyl bond —CH₂— 4-CH₂—N⁺(CH₃)₂-cyclohexyl, 3 3-CF₃-phenyl bond—CH₂— 4-CH₂—N⁺(CH₃)₂-cyclohexyl, 4 3,4-Cl₂-phenyl —CH═CH— —CH₂—4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 5 3-Br-phenyl —CH═CH— —CH₂—4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 6 phenyl bond bond4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 7 3,4-Cl₂-phenyl bond bond3-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 8 3-Br-phenyl bond bond3-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 9 2,3-Cl₂-phenyl bond bond4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 10 2,4-Cl₂-phenyl bond bond4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 11 2,5-Cl₂-phenyl bond bond4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 12 2,6-Cl₂-phenyl bond bond4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 13 2-Cl-phenyl bond bond4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 14 3,4-Cl₂-phenyl bond bond4-CH₂—N⁺(CH₃)₂-bicyclo[2.2.1]hept-2- yl, 15 3,4-Cl₂-phenyl bond bond4-CH₂—N⁺(CH₃)₂-(2S)-CH₂-tetrahydro- furan-2-yl, 16 3,4-Cl₂-phenyl bondbond 4-CH₂—N⁺(CH₃)₂-(2R)-CH₂-tetrahydro- furan-2-yl, 17 3,4-Cl₂-phenylbond bond 4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 18 3,4-Cl₂-phenyl bondbond 4-CH₂—N⁺(CH₃)₂-CH₂-tetrahydro-pyran- 4-yl, 19 3,4-Cl₂-phenyl bondbond 4-CH₂—N⁺(CH₃)₂-tetrahydro-thien-3-yl, 20 3,4-Cl₂-phenyl bond bond4-CH₂—N⁺(CH₃)₂-tetrahydro-thiopyran- 4-yl, 21 3,4-Cl₂-phenyl bond bond4-CH₂—N⁺[(CH₃)(CH₂CH₃)]-tetrahydro- pyran-4-yl, 22 3,4-Cl₂-phenyl bondbond 4-CH₂—N⁺{(CH₃)[(CH₂)₂CH₃)]}- tetrahydro-pyran-4-yl, 233,5-Cl₂-phenyl bond bond 4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 243-Br-phenyl bond bond 4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 252-CH₃-3-Cl₂-phenyl bond bond 4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 263-Cl-4-F-phenyl bond bond 4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 273-Cl-4-OCH₃-phenyl bond bond 4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 283-Cl-4-CH₃-phenyl bond bond 4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 293-Cl-phenyl bond bond 4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 303-CN-phenyl bond bond 4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 313-OCH₃-phenyl bond bond 4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 322-CH₃-4-Cl-phenyl bond bond 4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 333-CF₃-4-Cl-phenyl bond bond 4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 344-Cl-phenyl bond bond 4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 352-CH₃-5-Cl-phenyl bond bond 4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 363,4-Cl₂-phenyl bond bond 4-(CH₂)₂—N⁺(CH₃)₂-tetrahydro-pyran-4- yl, 373-Br-phenyl bond bond 4-(CH₂)₂—N⁺(CH₃)₂-tetrahydro-pyran-4- yl, 383-Br-phenyl —CH═CH— bond 3-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 393,4-Cl₂-phenyl bond —CH₂— 4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 403,4-Cl₂-phenyl —CH═CH— bond 4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 413,4-Cl₂-phenyl —CH═CH— bond 4-CH₂—N⁺(CH₃)₂-tetrahydro-thiopyran- 4-yl,42 3,5-F₂-phenyl —CH═CH— bond 4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 433-Br-phenyl —CH═CH— bond 4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 443-Br-phenyl —CH═CH— bond 4-CH₂—N⁺(CH₃)₂-tetrahydro-thiopyran- 4-yl, 453-Cl-phenyl —CH═CH— bond 4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 463-F-phenyl —CH═CH— bond 4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 474-Br-phenyl —CH═CH— bond 4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 483,4-Cl₂-phenyl —CH═CH— —CH₂— 4-CH₂-(1-CH₃-piperidinium), 49 3-Br-phenyl—CH═CH— —CH₂— 4-CH₂-(1-CH₃-piperidinium), 50 3,4-Cl₂-phenyl bond bond4-CH₂-(1-CH₃-piperidinium), 51 3,4-Cl₂-phenyl bond bond4-CH₂-(1-CH₃-pyrrolidinium), 52 3-Br-phenyl —CH═CH— bond3-CH₂-(1-CH₃-piperidinium), 53 3,4-Cl₂-phenyl —CH═CH— bond4-CH₂-(1-CH₃-piperidinium), 54 3,4-Cl₂-phenyl —CH═CH— bond4-CH₂-[4-(2-OCH₃-phenyl)-1-CH₃- piperazin-1-ium], 55 3-Br-phenyl —CH═CH—bond 4-CH₂-(1-CH₃-piperidinium), 56 3-CF₃-phenyl bond bond3-CH₂-(1-CH₃-piperidinium), 57 3-CF₃-phenyl —CH═CH— bond4-CH₂-(1-CH₃-piperidinium), 58 3,4-Cl₂-phenyl —CH═CH— —CH₂—4-CH₂-(4-CH₃-morpholin-4-ium), 59 3,4-Cl₂-phenyl bond bond4-CH₂-(4-CH₃-morpholin-4-ium), 60 3,4-Cl₂-phenyl —CH═CH— bond4-CH₂-(4-CH₃-morpholin-4-ium), 61 3-Br-phenyl —CH═CH— bond4-CH₂-(4-CH₃-morpholin-4-ium), 62 3-CF₃-phenyl —CH═CH— —CH₂—4-CH₂-(4-CH₃-morpholin-4-ium), 63 3-Br-phenyl —CH═CH— bond4-CH₂—N⁺[(CH₃)(CH₂CH═CH₂)]- tetrahydro-thiopyran-4-yl, 64 3-CF₃-phenyl—CH═CH— —CH₂— 4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 65 3-CF₃-phenyl bondbond 3-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 66 3-CH₃-phenyl —CH═CH— bond4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 67 3-CF₃-phenyl bond bond4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 68 3-CF₃-phenyl —CH═CH— bond4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 69 3-CH₃-phenyl bond bond4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 70 3,4-Cl₂-phenyl bond bond4-CH₂—N⁺(CH₃)₂-cycloheptyl, 71 3,4-Cl₂-phenyl —CH═CH— bond4-CH₂—N⁺(CH₃)₂-cyclohexyl, 72 3-Br-phenyl —CH═CH— bond4-CH₂—N⁺(CH₃)₂-cyclohexyl, 73 3-Br-phenyl bond bond4-CH₂—N⁺(CH₃)₂-cyclohexyl, 74 3-CF₃-phenyl bond bond4-CH₂—N⁺(CH₃)₂-cyclohexyl, 75 3,4-Cl₂-phenyl bond bond4-CH₂—N⁺(CH₃)₂-cyclohexyl, 76 3-Cl-4-F-phenyl bond bond4-CH₂—N⁺(CH₃)₂-cyclohexyl, 77 2,3-Cl₂-phenyl bond bond4-CH₂—N⁺(CH₃)₂-cyclohexyl, 78 2,6-Cl₂-phenyl bond bond4-CH₂—N⁺(CH₃)₂-cyclohexyl, 79 3-Cl-4-OCH₃-phenyl bond bond4-CH₂—N⁺(CH₃)₂-cyclohexyl, 80 3-Cl-4-CH₃-phenyl bond bond4-CH₂—N⁺(CH₃)₂-cyclohexyl, 81 2,5-Cl₂-phenyl bond bond4-CH₂—N⁺(CH₃)₂-cyclohexyl, 82 3,4-Cl₂-phenyl bond bond4-CH₂—N⁺(CH₃)₂-cyclopentyl, 83 3,4-Cl₂-phenyl —CH═CH— bond3-CH₂—N⁺(CH₃)₂-cyclohexyl, 84 4-F-phenyl —CH═CH— bond3-CH₂—N⁺(CH₃)₂-cyclohexyl, 85 3-(4-CF₃-phenyl)- bond bond4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, phenyl 86 3-(4-CH₃-phenyl)- bondbond 4-CH₂—N⁺(CH₃)₂-cyclohexyl, phenyl 87 3-(4-CH₃-phenyl)- bond bond4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, phenyl 88 4-biphenyl bond bond4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 89 1-naphthalene bond bond4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 90 2-naphthalene bond bond4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 91 2-naphthalene bond bond4-CH₂—N⁺[(CH₃)(CH₂CH₃)]-tetrahydro- pyran-4-yl, 92 2-naphthalene bondbond 4-CH₂—N⁺{(CH₃)[(CH₂)₂CH₃)]}- tetrahydro-pyran-4-yl, 937-Br-naphthalen-2- bond bond 4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, yl 947-Br-naphthalen-2- bond bond 4-CH₂—N⁺(CH₃)₂-cyclohexyl, yl 956-Br-2H-chromen-3- bond bond 4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, yl 966-Cl-2H-chromen-3- bond bond 4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, yl 976-Br-2H-chromen-3- bond bond 4-CH₂—N⁺(CH₃)₂-cyclohexyl, yl 986-Cl-2H-chromen-3- bond bond 4-CH₂—N⁺(CH₃)₂-cyclohexyl, yl 996-Br-2H-chromen-3- bond —CH₂— 4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, yl100 5,7-Cl₂-2H- bond bond 4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl,chromen-3-yl 101 5,7-Cl₂-2H- bond bond 4-CH₂—N⁺(CH₃)₂-cyclohexyl,chromen-3-yl 102 6,8-Cl₂-2H- bond bond4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, chromen-3-yl 103 6-CH₃-2H-chromen-bond bond 4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 3-yl 104 6-OCH₃-2H- bondbond 4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, chromen-3-yl 1056-CH₃-2H-chromen- bond bond 4-CH₂—N⁺(CH₃)₂-cyclohexyl, 3-yl 1066-OCH₃-2H- bond bond 4-CH₂—N⁺(CH₃)₂-cyclohexyl, chromen-3-yl 1076,8-Cl₂-2H- bond bond 4-CH₂—N⁺(CH₃)₂-cyclohexyl, chromen-3-yl 1086-Cl-2H-chromen-3- bond bond 4-CH₂—N⁺(CH₃)₂-(2R)-CH₂-tetrahydro- ylfuran-2-yl, 109 6-Cl-2H-chromen-3- bond bond4-CH₂—N⁺(CH₃)₂-(2S)-CH₂-tetrahydro- yl furan-2-yl, 1106-Cl-2H-chromen-3- bond bond 4-CH₂—N⁺(CH₃)₂-(2S)- ylbicyclo[2.2.1]hept-2-yl, 111 6,8-Cl₂-2H- bond bond4-CH₂—N⁺(CH₃)₂-bicyclo[2.2.1]hept-2- chromen-3-yl yl, 1128-CH₃-2H-chromen- bond bond 4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 3-yl113 8-CH₃-2H-chromen- bond bond 4-CH₂—N⁺(CH₃)₂-cyclohexyl, 3-yl 1146-Cl-8-CH₃-2H- bond bond 4-CH₂—N⁺(CH₃)₂-cyclohexyl, chromen-3-yl 1156-Cl-8-CH₃-2H- bond bond 4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl,chromen-3-yl 116 7,8-Cl₂-2H- bond bond 4-CH₂—N⁺(CH₃)₂-cyclohexyl,chromen-3-yl 117 6-Cl-8-CH₃-2H- bond bond4-CH₂—N⁺(CH₃)₂-bicyclo[2.2.1]hept-2- chromen-3-yl yl, 118 6-Cl-8-CH₃-2H-bond bond 4-CH₂—N⁺(CH₃)₂-cycloheptyl, chromen-3-yl 119 6-Cl-8-CH₃-2H-bond bond 4-CH₂—N⁺(CH₃)₂-cyclopentyl, chromen-3-yl 120 6-Cl-8-CH₃-2H-bond bond 4-CH₂—N⁺(CH₃)₂-thien-3-yl, chromen-3-yl 121 6-Cl-8-CH₃-2H-bond —CH₂— 4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, chromen-3-yl 1226,8-Cl₂-2H- bond bond 4-CH₂—N⁺(CH₃)₂-thien-3-yl, chromen-3-yl 1236-F-2H-chromen-3- bond bond 4-CH₂—N⁺(CH₃)₂-cyclohexyl, yl 1245-F-2H-chromen-3- bond bond 4-CH₂—N⁺(CH₃)₂-cyclohexyl, yl 1256-CF₃-2H-chromen- bond bond 4-CH₂—N⁺(CH₃)₂-cyclohexyl, 3-yl 1268-F-2H-chromen-3- bond bond 4-CH₂—N⁺(CH₃)₂-cyclohexyl, yl 1277-CH₃-2H-chromen- bond bond 4-CH₂—N⁺(CH₃)₂-cyclohexyl, 3-yl 1287-OCH₃-2H- bond bond 4-CH₂—N⁺(CH₃)₂-cyclohexyl, chromen-3-yl 1296-OCH₃-2H- bond bond 4-CH₂—N⁺(CH₃)₂-cyclohexyl, chromen-3-yl 1306-CF₃-2H-chromen- bond bond 4-CH₂—N⁺(CH₃)₂-thien-3-yl, 3-yl 1314-F-2H-chromen-3- bond bond 4-CH₂—N⁺(CH₃)₂-thien-3-yl, yl 1325-F-2H-chromen-3- bond bond 4-CH₂—N⁺(CH₃)₂-thien-3-yl, yl 1334-CF₃-2H-chromen- bond bond 4-CH₂—N⁺(CH₃)₂-cyclohexyl, 3-yl 1348-CF₃-2H-chromen- bond bond 4-CH₂—N⁺(CH₃)₂-cyclohexyl, 3-yl 135 3H- bondbond 4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, benzo[f]chromen-2- yl 136 3H-bond bond 4-CH₂-(1-CH₃-pyrrolidinium), benzo[f]chromen-2- yl 137 3H-bond bond 4-CH₂—N⁺(CH₃)₂-cyclohexyl, benzo[f]chromen-2- yl 138 3H- bondbond 4-CH₂—N⁺(CH₃)₂-tetrahydro-thiopyran- benzo[f]chromen-2- 4-yl, yl139 3H- bond bond 4-CH₂-(4-CH₃-morpholin-4-ium), benzo[f]chromen-2- yl140 3H- bond bond 4-CH₂—N⁺(CH₃)₂-CH₂-tetrahydro-pyran-benzo[f]chromen-2- 4-yl, yl 141 3H- bond —CH₂—4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, benzo[f]chromen-2- yl 1423-Br-8,9-dihydro- bond —CH₂— 4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 7H-benzocyclohepten-6- yl 143 3-Br-8,9-dihydro- bond bond4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 7H- benzocyclohepten-6- yl 1443-Br-8,9-dihydro- bond bond 4-CH₂—N⁺(CH₃)₂-cyclohexyl, 7H-benzocyclohepten-6- yl 145 8,9-dihydro-7H- bond bond4-CH₂—(1-CH₃-pyrrolidinium), benzocyclohepten-6- yl 146 8,9-dihydro-7H-bond bond 4-CH₂—N⁺(CH₃)₂-cyclohexyl, benzocyclohepten-6- yl 1478,9-dihydro-7H- bond bond 4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl,benzocyclohepten-6- yl 148 8,9-dihydro-7H- bond —CH₂—4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, benzocyclohepten-6- yl 149(2-CH₃-5-phenyl)- bond —CH₂— 4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl,furan-3-yl 150 [5-(4-Cl-phenyl)-2- bond —CH₂—4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, CH₃]-furan-3-yl 151(2-CH₃-5-phenyl)- bond bond 4-CH₂—N⁺(CH₃)₂-cyclohexyl, furan-3-yl 152benzofuran-2-yl bond bond 4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 153[5-(4-Cl-phenyl)-2- bond —CH₂— 4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl,CF₃]-furan-3-yl 154 [5-(4-Cl-phenyl)-2- bond —CH₂—4-CH₂—N⁺(CH₃)₂-cyclohexyl, CF₃]-furan-3-yl 155 5-Cl-benzofuran-2- bondbond 4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, yl 156 5-Cl-benzofuran-2-bond bond 4-CH₂—N⁺(CH₃)₂-cyclohexyl, yl 157 benzofuran-2-yl bond bond4-CH₂—N⁺(CH₃)₂-cyclohexyl, 158 1-CH₃-1H-indol-2-yl bond bond4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 159 5-Cl-1H-indol-2-yl bond bond4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 160 5-Br-1H-indol-2-yl bond bond4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 161 1-CH₃-1H-indol-3-yl bond bond4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 162 (1-CH₂-phenyl)-1H- bond bond4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, indol-3-yl 163 1-CH₃-1H-indol-2-ylbond bond 4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 164 5-Cl-1H-indol-2-ylbond bond 4-CH₂—N⁺(CH₃)₂-cyclohexyl, 165 5-Cl-1H-indol-2-yl bond bond4-CH₂—N⁺(CH₃)₂-(2S)-CH₂-tetrahydro- furan-2-yl, 166 5-Cl-1H-indol-2-ylbond bond 4-CH₂—N⁺(CH₃)₂—CH₂— bicyclo[2.2.1]hept-2-yl, 1677,8-Cl₂-2,3-dihydro- bond bond 4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl,benzo[b]oxepin-4-yl 168 7,8-Cl₂-2,3-dihydro- bond bond4-CH₂—N⁺(CH₃)₂-cyclohexyl, benzo[b]oxepin-4-yl 169 7,8-Cl₂-2,3-dihydro-bond bond 4-CH₂—N⁺(CH₃)₂-bicyclo[2.2.1]hept-2- benzo[b]oxepin-4-yl yl,170 7,8-Cl₂-2,3-dihydro- bond —CH₂—4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, benzo[b]oxepin-4-yl 1717,8-Cl₂-2,3-dihydro- bond bond 4-CH₂—N⁺(CH₃)₂-thien-3-yl,benzo[b]oxepin-4-yl 172 5-Br-pyridin-3-yl bond bond4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 173 2-Cl-pyridin-4-yl bond bond4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 174 3-Cl-benzo[b]thien- bond bond4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 2-yl 175 2,5-Cl₂-thien-3-yl bondbond 4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 176 benzo[b]thien-2-yl bondbond 4-CH₂—N⁺(CH₃)₂-tetrahydro-pyran-4-yl, 177 benzo[b]thien-2-yl bondbond 4-CH₂—N⁺(CH₃)₂-cyclohexyl, 178 3-Cl-benzo[b]thien- bond bond4-CH₂—N⁺(CH₃)₂-cyclohexyl, 2-yl

An example of the invention is a compound of Formula (I) andpharmaceutically acceptable forms thereof represented as follows:

An example of the invention is a compound of Formula (I) andpharmaceutically acceptable forms thereof selected from:

Definitions

Bond lines drawn into a ring system from a substituent variable indicatethat the substituent may be attached to any of the substitutable ringatoms.

As used herein, the following terms are intended to have the followingdefinitions.

The term “alkyl” means a saturated aliphatic branched or straight-chainmonovalent hydrocarbon radical or linking group substituent having from1-8 carbon atoms, wherein the radical is derived by the removal of onehydrogen atom from a carbon atom and the linking group is derived by theremoval of one hydrogen atom from each of two carbon atoms in the chain.The term includes, without limitation, methyl, ethyl, propyl, butyl,pentyl, hexyl and the like. An alkyl substituent may be attached to acore molecule via a terminal carbon atom or via a carbon atom within thechain. Similarly, any number of substituent variables may be attached toan alkyl substituent when allowed by available valences. The term “loweralkyl” means an alkyl substituent having from 1-4 carbon atoms.

The term “alkenyl” means a partially unsaturated alkyl substituenthaving at least one double bond derived by the removal of one hydrogenatom from each of two adjacent carbon atoms in the chain. The termincludes, without limitation, vinyl, vinylidene, allyl, allylidene,isopropenyl, prenyl, methallyl and the like. An alkenyl substituent maybe attached to a core molecule via a terminal carbon atom or via acarbon atom within the chain. Similarly, any number of substituentvariables may be attached to an alkenyl substituent when allowed byavailable valences. The term “lower alkenyl” means an alkenylsubstituent having from 1-4 carbon atoms.

The term “alkoxy” means an alkyl radical or linking group substituentattached through an oxygen-linking atom. The term includes, withoutlimitation, methoxy, ethoxy, propoxy, butoxy and the like. An alkoxysubstituent may be attached to a core molecule and further substitutedwhere allowed.

The term “cycloalkyl” means a monovalent saturated or partiallyunsaturated monocyclic, polycyclic or bridged hydrocarbon ring systemradical or linking group substituent. A ring of 3 to 20 carbon atoms maybe designated by C₃₋₂₀ cycloalkyl; a ring of 5 to 15 carbon atoms may bedesignated by C₅₋₁₅ cycloalkyl; a ring of 3 to 8 carbon atoms may bedesignated by C₃₋₈ cycloalkyl and the like. The term includes, withoutlimitation, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,cycloheptyl, cyclooctyl, 1H-indenyl, indanyl,1,2,3,4-tetrahydro-naphthalenyl, 5,6,7,8-tetrahydro-naphthalenyl,8,9-dihydro-7H-benzocyclohepten-6-yl,6,7,8,9-tetrahydro-5H-benzocycloheptenyl,5,6,7,8,9,10-hexahydro-benzocyclooctenyl, fluorenyl,bicyclo[2.2.1]heptyl, bicyclo[2.2.1]heptenyl, bicyclo[2.2.2]octyl,bicyclo[3.1.1]heptyl, bicyclo[3.2.1]octyl, bicyclo[2.2.2]octenyl,bicyclo[3.2.1]octenyl, adamantanyl, octahydro-4,7-methano-1H-indenyl,octahydro-2,5-methano-pentalenyl and the like. A cycloalkyl substituentmay be attached to a core molecule and further substituted whereallowed.

The term “aryl” means an unsaturated, conjugated π electron monocyclicor polycyclic hydrocarbon ring system radical or linking groupsubstituent of 6, 9, 10 or 14 carbon atoms.

The term includes, without limitation, phenyl, naphthalenyl, fluorenyl,indenyl, azulenyl, anthracenyl and the like. An aryl substituent may beattached to a core molecule and further substituted where allowed.

The term “heterocyclyl” means a saturated, partially unsaturated (suchas those named with the prefix dihydro, trihydro, tetrahydro, hexahydroand the like) or unsaturated monocyclic, polycyclic or bridgedhydrocarbon ring system radical or linking group substituent, wherein atleast one ring carbon atom has been replaced with one or moreheteroatoms independently selected from N, O or S. A heterocyclylsubstituent further includes a ring system having up to 4 nitrogen atomring members or a ring system having from 0 to 3 nitrogen atom ringmembers and 1 oxygen or sulfur atom ring member. Alternatively, up totwo adjacent ring members may be a heteroatom, wherein one heteroatom isnitrogen and the other is selected from N, O or S. A heterocyclylradical is derived by the removal of one hydrogen atom from a singlecarbon or nitrogen ring atom. A heterocyclyl linking group is derived bythe removal of one hydrogen atom from two of either a carbon or nitrogenring atom. A heterocyclyl substituent may be attached to a core moleculeby either a carbon atom ring member or by a nitrogen atom ring memberand further substituted where allowed.

The term heterocyclyl includes, without limitation, furanyl, thienyl,2H-pyrrolyl, 2-pyrrolinyl, 3-pyrrolinyl, pyrrolidinyl, pyrrolyl,1,3-dioxolanyl, oxazolyl, thiazolyl, imidazolyl, 2-imidazolinyl (alsoreferred to as 4,5-dihydro-1H-imidazolyl), imidazolidinyl,2-pyrazolinyl, pyrazolidinyl, pyrazolyl, isoxazolyl, isothiazolyl,oxadiazolyl, triazolyl, thiadiazglyl, tetrazolyl, tetrazolinyl,tetrazolidinyl, 2H-pyranyl, 4H-pyranyl, thiopyranyl, pyridinyl,piperidinyl, 1,4-dioxanyl, morpholinyl, 1,4-dithianyl, thiomorpholinyl,pyridazinyl, pyrimidinyl, pyrazinyl, piperazinyl, azetidinyl, azepanyl,indolizinyl, indolyl, isoindolyl, 3H-indolyl, indolinyl, benzofuranyl,benzo[b]thienyl, 1H-indazolyl, benzoimidazolyl, benzothiazolyl, purinyl,4H-quinolizinyl, quinolinyl, isoquinolinyl, cinnolinyl, phthalzinyl,quinazolinyl, quinoxalinyl, 1,8-naphthyridinyl, pteridinyl,quinuclidinyl, 2H-chromenyl, 3H-benzo[f]chromenyl, tetrahydro-furanyl,tetrahydro-thienyl, tetrahydro-pyranyl, tetrahydro-thiopyranyl,tetrahydro-pyridazinyl, hexahydro-1,4-diazepinyl,hexahydro-1,4-oxazepanyl, 2,3-dihydro-benzo[b]oxepinyl,1,3-benzodioxolyl (also known as 1,3-methylenedioxyphenyl),2,3-dihydro-1,4-benzodioxinyl (also known as 1,4-ethylenedioxyphenyl),benzo-dihydro-furanyl (also known as 2,3-dihydro-benzofuranyl),benzo-tetrahydro-pyranyl, benzo-dihydro-thienyl,5,6,7,8-tetrahydro-4H-cyclohepta[b]thienyl;5,6,7-trihydro-4H-cyclohexa[b]thienyl,5,6-dihydro-4H-cyclopenta[b]thienyl, 2-aza-bicyclo[2.2.1]heptyl,1-aza-bicyclo[2.2.2]octyl, 8-aza-bicyclo[3.2.1]octyl,7-oxa-bicyclo[2.2.1]heptyl, pyrrolidinium, piperidinium, piperazinium,morpholinium and the like.

The term “independently selected” refers to two or more substituentsthat may be selected from a substituent variable group, wherein theselected substituents may be the same or different.

The term “dependently selected” refers to one or more substituentvariables that are specified in an indicated combination forsubstitution in a core molecule (e.g., variables that refer to groups ofsubstituents appearing in a tabular list of compounds).

The term “carbonyl” means a linking group having the formula —C(O)— or—C(═O)—.

The term “thiocarbonyl” means a linking group having the formula —C(S)—or —C(═S)—.

The term “sulfonyl” means a linking group having the formula —SO₂—.

The term “alkoxycarbonyl” means a radical having the formula—C(O)O-alkyl.

Pharmaceutically Acceptable Forms

Pharmaceutically acceptable forms according to the invention may,alternatively or in addition to a compound of Formula (I), comprise apharmaceutically acceptable salt of a compound of Formula (I) or aprodrug or pharmaceutically active metabolite of such a compound orsalt.

The compounds of the invention may be present in the form ofpharmaceutically acceptable salts. For use in medicines, the salts ofthe compounds of this invention refer to non-toxic “pharmaceuticallyacceptable salts.” FDA-approved pharmaceutically acceptable salt formsinclude pharmaceutically acceptable acidic/anionic or basic/cationicsalts.

Pharmaceutically acceptable acidic/anionic salts include, withoutlimitation, acetate, benzenesulfonate, benzoate, bicarbonate,bitartrate, bromide, calcium edetate, camsylate, carbonate, chloride,citrate, dihydrochloride, edetate, edisylate, estolate, esylate,fumarate, glyceptate, gluconate, glutamate, glycollylarsanilate,hexylresorcinate, hydrabamine, hydrobromide, hydrochloride,hydroxynaphthoate, iodide, isethionate, lactate, lactobionate, malate,maleate, mandelate, mesylate, methylbromide, methylnitrate,methylsulfate, mucate, napsylate, nitrate, pamoate, pantothenate,phosphate/diphosphate, polygalacturonate, salicylate, stearate,subacetate, succinate, sulfate, tannate, tartrate, teoclate, tosylateand triethiodide salts.

Organic or inorganic acids also include, and are not limited to,hydroiodic, perchloric, sulfuric, phosphoric, propionic, glycolic,methanesulfonic, hydroxyethanesulfonic, oxalic, 2-naphthalenesulfonic,p-toluenesulfonic, cyclohexanesulfamic, saccharinic or trifluoroaceticacid.

Pharmaceutically acceptable basic/cationic salts include, and are notlimited to aluminum, 2-amino-2-hydroxymethyl-propane-1,3-diol (alsoknown as tris(hydroxymethyl)aminomethane, tromethane or “TRIS”),ammonia, benzathine, t-butylamine, calcium, calcium gluconate, calciumhydroxide, chloroprocaine, choline, choline bicarbonate, cholinechloride, cyclohexylamine, diethanolamine, ethylenediamine, lithium,LiOMe, L-lysine, magnesium, meglumine, NH₃, NH₄OH, N-methyl-D-glucamine,piperidine, potassium, potassium-t-butoxide, potassium hydroxide(aqueous), procaine, quinine, sodium, sodium carbonate,sodium-2-ethylhexanoate (SEH), sodium hydroxide, triethanolamine (TEA)or zinc.

The compounds of the invention may be present in the form ofpharmaceutically acceptable prodrugs and metabolites thereof. Ingeneral, such prodrugs and metabolites will be functional derivatives ofthe compounds that are readily convertible in vivo into an activecompound.

The term “prodrug” means a pharmaceutically acceptable form of afunctional derivative of a compound of the invention (or a saltthereof), wherein the prodrug may be: 1) a relatively active precursorwhich converts in vivo to an active prodrug component; 2) a relativelyinactive precursor which converts' in vivo to an active prodrugcomponent; or 3) a relatively less active component of the compound thatcontributes to therapeutic biological activity after becoming availablein vivo (i.e., as a metabolite). Conventional procedures for theselection and preparation of suitable prodrug derivatives are describedin, for example, “Design of Prodrugs”, ed. H. Bundgaard, Elsevier, 1985.

The term “metabolite” means a pharmaceutically acceptable form of ametabolic derivative of a compound of the invention (or a salt thereof),wherein the derivative is a relatively less active component of thecompound that contributes to therapeutic biological activity afterbecoming available in vivo.

The present invention also contemplates compounds of Formula (I) invarious stereoisomeric or tautomeric forms. The invention encompassesall such CCR2 inhibiting compounds, including active compounds in theform of essentially pure enantiomers, racemic mixtures and tautomers orpharmaceutically acceptable forms thereof.

The term “isomer” refers to compounds that have the same composition andmolecular weight but differ in physical and/or chemical properties. Suchsubstances have the same number and kind of atoms but differ instructure. The structural difference may be in constitution (geometricisomers) or in an ability to rotate the plane of polarized light(stereoisomers).

The term “stereoisomer” refers to isomers of identical constitution thatdiffer in the arrangement of their atoms in space. Enantiomers anddiastereomers are stereoisomers wherein an asymmetrically substitutedcarbon atom acts as a chiral center. The term “chiral” refers to amolecule that is not superposable on its mirror image, implying theabsence of an axis and a plane or center of symmetry. The term“enantiomer” refers to one of a pair of molecular species that aremirror images of each other and are not superposable. The term“diastereomer” refers to stereoisomers that are not related as mirrorimages. The symbols “R” and “S” represent the configuration ofsubstituents around a chiral carbon atom(s). The symbols “R*” and “S*”denote the relative configurations of substituents around a chiralcarbon atom(s).

The term “racemate” or “racemic mixture” refers to a compound ofequimolar quantities of two enantiomeric species, wherein the compoundis devoid of optical activity. The term “optical activity” refers to thedegree to which a chiral molecule or nonracemic mixture of chiralmolecules rotates the plane of polarized light.

The term “geometric isomer” refers to isomers that differ in theorientation of substituent atoms in relationship to a carbon-carbondouble bond, to a cycloalkyl ring or to a bridged bicyclic system.Substituent atoms (other than H) on each side of a carbon-carbon doublebond may be in an E or Z configuration. In the “E” or “chair”configuration, the substituents are on opposite sides in relationship tothe carbon-carbon double bond; in the “Z” or “boat” configuration, thesubstituents are oriented on the same side in relationship to thecarbon-carbon double bond.

Substituent atoms (other than H) attached to a hydrocarbon ring may bein a cis or trans configuration. In the “cis” configuration, thesubstituents are on the same side in relationship to the plane of thering; in the “trans” configuration, the substituents are on oppositesides in relationship to the plane of the ring. Compounds having amixture of “cis” and “trans” species are designated “cis/trans”.Substituent atoms (other than H) attached to a bridged bicyclic systemmay be in an “endo” or “exo” configuration. In the “endo” configuration,the substituents attached to a bridge (not a bridgehead) point towardthe larger of the two remaining bridges; in the “exo” configuration, thesubstituents attached to a bridge point toward the smaller of the tworemaining bridges.

It is to be understood that the various substituent stereoisomers,geometric isomers and mixtures thereof used to prepare compounds of thepresent invention are either commercially available, can be preparedsynthetically from commercially available starting materials or can beprepared as isomeric mixtures and then obtained as resolved isomersusing techniques well-known to those of ordinary skill in the art.

The isomeric descriptors “R,” “S,” “S*,” “R*,” “E,” “Z,” “cis,” “trans”,“exo”, and “endo”, where used herein, indicate atom configurationsrelative to a core molecule and are intended to be used as defined inthe literature.

The compounds of the present invention may be prepared as individualisomers by either isomer-specific synthesis or resolved from an isomericmixture. Conventional resolution techniques include forming the freebase of each isomer of an isomeric pair using an optically active salt(followed by fractional crystallization and regeneration of the freebase), forming an ester or amide of each of the isomers of an isomericpair (followed by chromatographic separation and removal of the chiralauxiliary) or resolving an isomeric mixture of either a startingmaterial or a final product using various well known chromatographicmethods.

Furthermore, compounds of the present invention may have a plurality ofpolymorph or amorphous crystalline forms and, as such, are intended tobe included in the scope of the invention. In addition, some of thecompounds may form a plurality of solvates with water (i.e., hydrates)or common organic solvents, such are also intended to be encompassedwithin the scope of this invention.

During any of the processes for preparation of the compounds of thepresent invention, it may be necessary and/or desirable to protectsensitive or reactive groups on any of the molecules concerned. This maybe achieved by means of conventional protecting groups, such as thosedescribed in Protective Groups in Organic Chemistry, ed. J. F. W.McOmie, Plenum Press, 1973; and T. W. Greene & P. G. M. Wuts, ProtectiveGroups in Organic Synthesis, John Wiley & Sons, 1991. The protectinggroups may be removed at a convenient subsequent stage using methodsknown in the art.

Therapeutic Use

Pharmaceutically acceptable forms of a compound of Formula (I) or acomposition or medicament thereof in accordance with the invention areCCR2 antagonists. Said composition or medicament may contain a compoundof Formula (I) having a mean inhibition constant (IC₅₀) against MCP-1binding to CCR2 of between about 5 μM to about 1 μM; between about 1 μMto about 1 nM; between about 800 nM to about 1 nM; between about 200 nMto about 1 nM; between about 100 nM to about 1 nM; between about 80 nMto about 1 nM; between about 20 nM to about 1 nM; between about 10 nM toabout 1 nM; or about 1 nM.

A compound of Formula (I) or a composition or medicament thereof reducesMCP-1 induced monocyte chemotaxis. Said composition or medicament maycontain a compound of Formula (I) having an IC₅₀ for reduction in MCP-1induced monocyte chemotaxis of between about 5 μM to about 1 nM; betweenabout 1 μM to about 1 nM; between about 800 nM to about 1 nM; betweenabout 200 nM to about 1 nM; between about 100 nM to about 1 nM; betweenabout 80 nM to about 1 nM; between about 20 nM to about 1 nM; betweenabout 10 nM to about 1 nM; or about 1 nM.

A compound of Formula (I) or a composition or medicament thereof reducesMCP-1 intracellular calcium mobilization. Said composition or medicamentmay contain a compound of Formula (I) having an IC₅₀ for reduction inMCP-1 induced intracellular calcium mobilization of between about 5 μMto about 1 nM; between about 1 μM to about 1 nM; between about 800 nM toabout 1 nM; between about 200 nM to about 1 nM; between about 100 nM toabout 1 nM; between about 80 nM to about 1 nM; between about 20 nM toabout 1 nM; between about 10 nM to about 1 nM; or about 0.1 nM.

Accordingly, a compound of Formula (I) or a composition or medicamentthereof is useful in a method for preventing, treating or ameliorating aCCR2 mediated inflammatory syndrome, disorder or disease in a subject inneed thereof comprising administering to the subject an effective amountof a compound of Formula (I) or composition or medicament thereof.

The present invention is directed to a method for preventing, treatingor ameliorating a CCR2 mediated inflammatory syndrome, disorder ordisease in a subject in need thereof comprising administering to thesubject an effective amount of a compound of Formula (I) or compositionor medicament thereof.

The term “administering,” with respect to the methods of the invention,means a method for preventing, treating or ameliorating a syndrome,disorder or disease as described herein with a compound of Formula (I)or composition or medicament thereof. Such methods include administeringan effective amount of said compound, composition or medicament atdifferent times during the course of a therapy or concurrently in acombination form. The methods of the invention are to be understood asembracing all known therapeutic treatment regimens.

The term “subject” as used herein, means an animal, typically a mammal,typically a human, typically a patient with a syndrome, disorder ordisease that is associated with elevated MCP-1 expression or MCP-1overexpression, or a patient with an inflammatory condition thataccompanies syndromes, disorders or diseases associated with elevatedMCP-1 expression or MCP-1 overexpression.

The term “effective amount” means that amount of active compound orpharmaceutical agent that elicits the biological or medicinal responsein a tissue system, animal or human, that is being sought by aresearcher, veterinarian, medical doctor, or other clinician, whichincludes preventing, treating or ameliorating the symptoms of asyndrome, disorder or disease being treated.

The effective amount of a compound of the invention in such atherapeutic method is from about 0.001 mg/kg/day to about 300 mg/kg/day.

The invention includes the use of an instant compound for thepreparation of a composition or medicament for preventing, treating orameliorating a CCR2 mediated inflammatory syndrome, disorder or diseasein a subject in need thereof, wherein the composition or medicamentcomprises a mixture one or more compounds of the invention and anoptional pharmaceutically acceptable carrier.

The term “composition” means a product comprising a compound of theinvention, such as a product comprising the specified ingredients in thespecified amounts, as well as any product which results, directly orindirectly, from such combinations of the specified ingredients in thespecified amounts and one or more pharmaceutically acceptable carrierstherefor.

The term “medicament” means a product for use in preventing, treating orameliorating a CCR2 mediated inflammatory syndrome, disorder or disease.

The term “pharmaceutically acceptable carrier” means molecular entitiesand compositions that are of sufficient purity and quality for use inthe formulation of a composition or medicament of the invention andthat, when appropriately administered to an animal or a human, do notproduce an adverse, allergic, of other untoward reaction. Since bothhuman and veterinary use is included within the scope of the invention,a pharmaceutically acceptable formulation includes a composition ormedicament for either human or veterinary use.

The term “CCR2 mediated inflammatory syndrome, disorder or disease”means, without limitation, syndromes, disorders or diseases associatedwith elevated MCP-1 expression, MCP-1 overexpression or inflammatoryconditions that accompany syndromes, disorders or diseases associatedwith elevated MCP-1 expression or MCP-1 overexpression.

The terms “elevated MCP-1 expression” or “MCP-1 overexpression” meanunregulated or up-regulated CCR2 activation as a result of MCP-1binding.

The term “unregulated” means unwanted CCR2 activation in a multicellularorganism resulting in harm (such as discomfort or decreased lifeexpectancy) to the multicellular organism.

The term “up-regulated” means: 1). increased or unregulated CCR2activity or expression, or 2). increased CCR2 expression leading tounwanted monocyte and lymphocyte migration. The existence of aninappropriate or abnormal level of MCP-1 or activity of CCR2 isdetermined by procedures well known in the art.

CCR2 mediated inflammatory syndromes, disorders or diseases include,without limitation, ophthalmic disorders, uveitis, atherosclerosis,rheumatoid arthritis, psoriasis, psoriatic arthritis, atopic dermatitis,multiple sclerosis, Crohn's Disease, ulcerative colitis, nephritis,organ allograft rejection, fibroid lung, renal insufficiency, diabetesand diabetic complications, diabetic nephropathy, diabetic retinopathy,diabetic retinitis, diabetic microangiopathy, tuberculosis, chronicobstructive pulmonary disease, sarcoidosis, invasive staphyloccocia,inflammation after cataract surgery, allergic rhinitis, allergicconjunctivitis, chronic urticaria, asthma, allergic asthma, periodontaldiseases, periodonitis, gingivitis, gum disease, diastoliccardiomyopathies, cardiac infarction, myocarditis, chronic heartfailure, angiostenosis, restenosis, reperfusion disorders,glomerulonephritis, solid tumors and cancers, chronic lymphocyticleukemia, chronic myelocytic leukemia, multiple myeloma, malignantmyeloma, Hodgkin's disease, and carcinomas of the bladder, breast,cervix, colon, lung, prostate, or stomach.

The term “uveitis” generically refers to any inflammatory diseaseinvolving the eye. Uveitis can be divided into clinically distinctsubtypes based on the part of the eye in which the inflammation ispresent (percentages correspond to patients known to fit thesecategories): anterior (51%), intermediate (13%), posterior (20%), orpanuveitis (16%) and, according to the course of the disease, as eitheracute (16%), recurring (26%), or chronic (58%). Those with 1.0 anterioruveitis (˜19%) eventually develop irreparable vision damage despiteaggressive treatment such as unilateral blindness (9%), bilateralblindness (2%), or unilateral or bilateral vision impairment (8%). Mostcases of uveitis are idiopathic, but known causes include infection(e.g., toxoplasmosis, cytomegalovirus, and the like) or development as acomponent of a systemic inflammatory and/or autoimmune disorder (e.g.,juvenile RA, HLA-B27-associated spondyloarthropathies, sarcoidosis, andthe like).

Patients with anterior uveitis have MCP-1 present in large quantities inthe aqueous humor of the eye. The amount of MCP-1 correlates with theseverity of the clinical symptoms and the large number of mononuclearcells present in the cellular infiltrate. Uveitis is also a potentialcomplication resulting from cataract surgery and prophylactic use ofantibiotics and corticosteroids is common for such patients. Currently,most patients with anterior uveitis are first treated with topicalcorticosteroids. Injected or oral steroids may be used in severe cases,or if the disease is recurrent or chronic. If steroids are ineffective,immunosuppressive agents (e.g., cyclosporine, methotrexate,azathioprine, cyclophosphamide, and the like) are used, particularly ifthe patient's vision is in danger. All of these drugs have potentiallysevere side-effects, particularly in children, and there is generalagreement that there is an unmet medical need for safe and effectivesteroid substitutes or steroid-sparing agents.

An example of the invention is a method for preventing, treating orameliorating CCR2 mediated ophthalmic disorders (such as uveitis,allergic conjunctivitis and the like), rheumatoid arthritis, psoriasis,psoriatic arthritis, atopic dermatitis, chronic obstructive pulmonarydisease, allergic rhinitis, asthma, allergic asthma, periodontaldiseases (such as periodonitis, gingivitis, gum disease and the like) ina subject in need thereof comprising administering to the subject aneffective amount of a compound of Formula (I) or composition ormedicament thereof.

Another example of the invention is a method for preventing, treating orameliorating CCR2 mediated uveitis, wherein uveitis includes, withoutlimitation, acute, recurring or chronic uveitis (such as anterioruveitis, intermediate uveitis, posterior uveitis, panuveitis and thelike) in a subject in need thereof comprising administering to thesubject an effective amount of a compound of Formula (I) or compositionor medicament thereof.

An example of the invention is a method for preventing, treating orameliorating CCR2 mediated acute uveitis, recurring uveitis, chronicuveitis, allergic conjunctivitis, rheumatoid arthritis, psoriasis,psoriatic arthritis, atopic dermatitis, chronic obstructive pulmonarydisease, allergic rhinitis, asthma, allergic asthma, periodonitis,gingivitis or gum disease in a subject in need thereof comprisingadministering to the subject an effective amount of a compound ofFormula (I) or composition or medicament thereof.

The invention includes a method for preventing, treating or amelioratinga CCR2 mediated inflammatory syndrome, disorder or disease in a subjectin need thereof comprising administering to the subject an effectiveamount of a compound of Formula (I) or composition or medicament thereofin a combination therapy with one or more anti-inflammatory agents (suchas a small molecule, antibiotic, corticosteroid, steroid, and the like),anti-infective agents or immunosuppressive agents.

The term “combination therapy” refers to the use of a compound ofFormula (I) or composition or medicament thereof in combination with ananti-inflammatory, anti-infective, or immunosuppressive agent forpreventing, treating or ameliorating a CCR2 mediated inflammatorysyndrome, disorder or disease.

For preventing, treating or ameliorating a CCR2 mediated inflammatorysyndrome, disorder or disease using a compound of Formula (I) orcomposition or medicament thereof and an anti-inflammatory,anti-infective or immunosuppressive agent in a combination therapyincludes, without limitation, co-administration of the compound and theagent, sequential administration of the compound and the agent,administration of a composition containing of the compound and the agentor simultaneous administration of separate compositions containing ofthe compound and the agent.

Pharmaceutical Compositions

The present invention includes a pharmaceutical composition ormedicament comprising one or more of the instant compounds and anoptional pharmaceutically acceptable carrier.

The present invention further includes a process for making apharmaceutical composition or medicament comprising mixing one or moreof the instant compounds and an optional pharmaceutically acceptablecarrier; and, includes those compositions or medicaments resulting fromsuch a process. Contemplated processes include both conventional andunconventional pharmaceutical techniques.

The composition or medicament may take a wide variety of forms toeffectuate mode of administration ocularly, intranasally (by inhalationor insufflation), sublingually, orally, parenterally or rectallyincluding, without limitation, ocular (via a delivery device such as acontact lens and the like), intranasal (via a delivery device),transdermal, topical with or without occlusion, intravenous (both bolusand infusion), injection (intraperitoneally, subcutaneously,intramuscularly, intratumorally, or parenterally) and the like.

The composition or medicament may be in a dosage unit such as a tablet,pill, capsule, powder, granule, liposome, biodegradable carrier, ionexchange resin, sterile solution and the like (facilitating immediaterelease, timed release, or sustained release), parenteral solution orsuspension, metered aerosol or liquid spray, drop, ampoule,auto-injector device or suppository.

Compositions or medicaments suitable for oral administration includesolid forms such as pills, tablets, caplets, capsules (each includingimmediate release, timed release, and sustained release formulations),granules and powders and liquid forms such as solutions, syrups,elixirs, emulsions and suspensions. Forms useful for nasaladministration include sterile solutions or nasal delivery devices.Forms useful for ocular administration include sterile solutions orocular delivery devices. Forms useful for parenteral administrationinclude sterile solutions, emulsions and suspensions.

Alternatively, the composition or medicament may be administered in aform suitable for once-weekly or once-monthly administration. Forexample, an insoluble salt of the active compound may be adapted toprovide a depot preparation for intramuscular injection (e.g., a saltform) or to provide a solution for nasal or ocular administration (e.g.,a quaternary ammonium salt).

The dosage form (tablet, capsule, powder, solution, contact lens, patch,liposome, ion exchange resin, suppository, teaspoonful, and the like)containing the composition or medicament thereof contains an effectiveamount of the active ingredient necessary to provide a therapeuticeffect.

The composition or medicament may contain an effective amount of fromabout 0.001 mg to about 5000 mg (preferably, from about 0.001 to about500 mg) of a compound of the present invention or a pharmaceuticallyacceptable form thereof and may be constituted into any form suitablefor the mode of administration selected for a subject in need.

A contemplated range of the effective amount includes from about 0.001mg to about 300 mg/kg of body weight per day. A contemplated range alsoincludes from about 0.003 to about 100 mg/kg of body weight per day.Another contemplated range includes from about 0.005 to about 15 mg/kgof body weight per day. The composition or medicament may beadministered according to a dosage Regimen of from about 1 to about 5times per day.

For oral administration, the composition or medicament is preferably inthe form of a tablet containing, e.g., 0.01, 0.05, 0.1, 0.5, 1.0, 2.5,5.0, 10.0, 15.0, 25.0, 50.0, 100, 150, 200, 250, and 500 milligrams ofthe active ingredient for the symptomatic adjustment of the dosage tothe patient to be treated. Optimal dosages will vary depending onfactors associated with the particular patient being treated (e.g., age,weight, diet, and time of administration), the severity of the conditionbeing treated, the compound being employed, the mode of administration,and the strength of the preparation. The use of either dailyadministration or post-periodic dosing may be employed.

For ocular administration, the composition is preferably in the form ofan ophthalmic composition. The ophthalmic compositions are preferablyformulated as eye-drop formulations and filled in appropriate containersto facilitate administration to the eye, for example a dropper fittedwith a suitable pipette. Preferably the compositions are sterile andaqueous based, using purified water. In addition to the compound of theinvention, an ophthalmic composition may contain one or more of: a) asurfactant such as a polyoxyethylene fatty acid ester; b) a thickeningagents such as cellulose, cellulose derivatives, carboxyvinyl polymers,polyvinyl polymers, and polyvinylpyrrolidones, typically at aconcentration n the range of about 0.05 to about 5.0% (wt/vol); c) (asan alternative to or in addition to storing the composition in acontainer containing nitrogen and optionally including a free oxygenabsorber such as Fe), an anti-oxidant such as butylated hydroxyanisol,ascorbic acid, sodium thiosulfate, or butylated hydroxytoluene at aconcentration of about 0.00005 to about 0.1% (wt/vol); d) ethanol at aconcentration of about 0.01 to 0.5% (wt/vol); and e) other excipientssuch as an isotonic agent, buffer, preservative, and/or pH-controllingagent. The pH of the ophthalmic composition is desirably within therange of 4 to 8.

Synthetic Methods

Representative compounds of the present invention can be synthesized inaccordance with the general synthetic schemes described below and areillustrated more particularly in the specific examples that follow. Thegeneral schemes and specific examples are offered by way ofillustration; the invention should not be construed as being limited bythe chemical reactions and conditions expressed. The methods forpreparing the various starting materials used in the schemes andexamples are well within the skill of persons versed in the art. Allcommercially available chemicals were obtained from commercial suppliersand used without further purification. Particular equipment componentsused in the examples such as reaction vessels and the like are alsocommercially available.

The following abbreviations have the indicated meanings:

Cpd compound Boc tert-butoxy carbonyl or t-butoxy carbonyl Boc₂Odi-t-butyl-dicarbonate conc. concentrated DABCO1,4-diazabicyclo[2.2.2]octane DBU 1,8-diazabicyclo[5.4.0]undec-7-ene DCMor CH₂Cl₂ dichloromethane DMAP (4,4-dimethylamino)-pyridine DMFN,N-dimethyl formamide EDIC1-(3-dimethylaminopropyl)-3-ethylcarbodiimide Et₂O ethoxy-ethane orether EtOAc ethylacetate EtOH ethanol HCOH formaldehyde HOBt1-hydroxybenzotriazole hydrate MeI or CH₃I methyl iodide MeOH methanolmin(s)/hr(s)/d(s) minute(s)/hour(s)/day(s) Pd(dppf)₂{[1,1′-bis(diphenylphosphino)-ferrocene] dichloropalladium(II)}RT/rt/r.t. room temperature NaB(OAc)₃H sodium triacetoxyborohydride TEAor Et₃N triethylamine THF tetrahydrofuran

Scheme A

The synthetic method provided by Scheme A, depending on the startingmaterials used or when certain reaction conditions are desired, is usedto prepare a compound of Formula (I).

A mixture of a nitrophenylamine hydrochloride Compound A1 and a solutionof an aldehyde or ketone Compound A2 (in a solvent such as CH₂Cl₂ andthe like) is cooled to 0° C., then Et₃N is added followed by NaB(OAc)₃H.The resulting suspension is stirred and allowed to warm to r.t. forabout 8-12 hrs (adapted from Shiroshi, et al., J. Med. Chem., 2000, 43,2049).

An aldehyde (such as formaldehyde and the like) in aqueous solution isadded, followed by NaB(OAc)₃H added in one portion while cooling thereaction vessel with ice. The reaction mixture is stirred at r.t. forabout 12 hrs, then made basic (using a solution of 2N NaOH, and thelike) and extracted (with a solvent such as CH₂Cl₂ and the like). Theorganic layer is washed using brine, then separated and dried overNa₂SO₄. The drying agent is filtered and the solvent is removed in vacuoto yield a disubstituted nitrophenylamine Compound A3 (adapted fromHashimoto et al., Org. Proc. R&D, 2002, 6, 70).

SnCl₂.2H₂O in conc. HCl is added in small portions to a solution ofCompound A3 (in a solvent such as THF and the like) at r.t. The mildlyexothermic reaction is maintained at r.t. using an ice-cooled water bathand stirred for about 1 hr. The round bottom flask containing thereaction mixture is placed in a warm water bath for about 30 min toallow the reaction to reach completion. The mixture is sequentiallydiluted with a solvent (such as THF and the like) and water, then madebasic (using a solution of 2N NaOH and the like). The layers areseparated and the aqueous layer is extracted (using a solvent such asEt₂O and the like). The combined organic layers were dried over MgSO₄,then filtered and the solvent was removed in vacuo. The solid obtainedwas further dried (by pressing the solid on an absorbent surface such asa filter paper) to provide a disubstituted aminophenylamine Compound A4.

A solution of a R₁ substituted Compound A5 (wherein Q represents aleaving group such as chloride or hydroxy) is added dropwise via adropping funnel over a period of about 20 min to a solution of CompoundA4 (in a solvent mixture such as Et₃N in THF and the like). Theresulting suspension is allowed to warm to r.t. over a period of about8-12 hrs, then made basic (using a solution of 2N NaOH and the like).The organic and aqueous layers are separately extracted (using a solventsuch as EtOAc and the like). The organic layer is washed with brine,then dried (using MgSO₄, Na₂SO₄ and the like) and filtered. The solventis removed in vacuo to yield a crude product which may be purified byeither flash column chromatography (in a solvent ratio 15:1 EtOAc:MeOHto 6:1 EtOAc:MeOH) or preparative TLC (using a solvent mixture in aratio of EtOAc:MeOH) to provide a substituted benzamide Compound A6.

An R₅ substituted iodo Compound A7 is added to a solution of Compound A6(in a solvent mixture such as acetone and acetonitrile and the like) atr.t. The resulting solution is stirred for about 8-12 hrs to form aprecipitate. The solvent is removed in vacuo and the solid issequentially washed (using EtOAc and Et₂O and the like), then dried in avacuum oven for about 12 hrs to provide a quaternary salt Compound A8.

A solution of Compound A8 (in a solvent:water ratio mix such ad a 1:1MeOH:H₂O) is passed through a column packed with an ion-exchange resin(such as ˜300 g of Bio-Rad analytical grade anion exchange resin. AG1-X8, 50-100 mesh, chloride form) into a flask. The column is thenwashed (such as with MeOH and the like) and solvents (such as acetoneand Et₂O and the like) are added to the filtered product in the flask:The solvent is removed in vacuo and the product dried in a vacuum ovenfor about 8-12 hrs to provide a target Compound A9 of Formula (I)wherein R₂ is N⁺(R₄R₅)—ZR₃ and pharmaceutically acceptable anionic saltforms thereof.

Scheme B

The synthetic method provided by Scheme B, depending on the startingmaterials used or when certain reaction conditions are desired, is usedas an alternative to Scheme A to prepare a compound of Formula (I).

Di-t-butyl-dicarbonate is added in one portion at r.t. to a solution ofCompound B1 (in a solvent such as CH₂Cl₂ and the like) and the solutionis stirred for about 48 hrs. The reaction mixture is sequentially washedwith an acidic solution (using a 10% citric acid solution and the like)and brine. The organic layer is separated, dried over Na₂SO₄ andfiltered. The solvent is removed in vacuo to provide a protectedCompound B2 that is used in the next step without further purification.A reagent (such as MnO₂ and the like) is added to a solution of CompoundB2 (in a solvent such as chloroform and the like) to form a blacksuspension, which is stirred at r.t. for about 8-12 hrs, then filteredthrough a pad of celite. The solvent was evaporated in vacuo to provideCompound B3, which is used in the next step without furtherpurification.

NaB(OAc)₃H is added to a mixture of Compound B3 and Compound B4 (in asolvent such as CH₂Cl₂ and the like) and the suspension is stirred atr.t. for a period of time. After the reaction is complete, an aldehyde(such as formaldehyde and the like) in aqueous solution is added,followed by NaB(OAc)₃H added in one portion while the reaction vessel iscooled with ice. The reaction mixture is stirred at r.t. for about 12hrs, then made basic (using a solution of 2N NaOH and the like) andextracted (with a solvent such as CH₂Cl₂ and the like). The organiclayer is washed using brine, then separated and dried over Na₂SO₄. Thedrying agent is filtered and the solvent is removed in vacuo to yield acrude product which was purified by column chromatography (using aneluent ratio of about 4:1 and a solvent mixture such as CH₂Cl₂:MeOH andthe like) to provide a disubstituted phenylamine Compound B5.

An HCl solution (in a solvent such as dioxane and the like) is added toa solution of Compound B5 (in a solvent such as CH₂Cl₂ and the like) andthe mixture is stirred at r.t. for about 12 hrs. The solvent is removedand the residue is made basic (using a base such as a 2N NaOH solution)then extracted (with a solvent such as EtOAc). The organic layer iswashed with brine, then separated and dried over Na₂SO₄. The dryingagent is filtered and the solvent is removed in vacuo to provide aCompound B6.

Accordingly, using the procedure of Scheme A to provide-additionalcompounds of the present invention, Compound B6 is used in place ofCompound A4 and carried forward to provide a target compound of Formula(I) wherein X₂ is —CH₂— and pharmaceutically acceptable anionic saltforms thereof.

Scheme C

The synthetic method provided by Scheme C, depending on the startingmaterials used or when certain reaction conditions are desired, is usedas an alternative to Scheme B to prepare a compound of Formula (I).

Using the procedure of Scheme B, Compound C1 is used in place ofCompound B4 and coupled with Compound B3 to provide Compound C2.

Accordingly, using the procedure of Scheme B to provide additionalcompounds of the present invention, Compound C2 is used in place ofCompound B5 and carried forward to provide a target compound of Formula(I) wherein X₂ is —CH₂— and R₄ and R₅ are taken together with thenitrogen atom of Formula-(I) to form a heterocyclyl ring andpharmaceutically acceptable anionic salt forms thereof.

Scheme D

The synthetic method provided by Scheme D, depending on the startingmaterials used or when certain reaction conditions are desired, is usedas an alternative to Scheme A or B to prepare a compound of Formula (I).

Using the procedure of Scheme B, Compound D1 and Compound B4 are coupledto provide Compound D2.

Accordingly, using the procedure of Scheme A to provide additionalcompounds of the present invention, Compound D2 is used in place ofCompound A3 and carried forward to provide a target compound of Formula(I) wherein X₂ is —CH₂— and pharmaceutically acceptable anionic saltforms thereof.

Scheme E

The synthetic method provided by Scheme E, depending on the startingmaterials used or when certain reaction conditions are desired, is usedto prepare a compound of Formula (I).

Compound E1 (wherein W₁ is a suitable leaving group such as a halogenatom) was reacted with Compound E2 to provide a further-substitutedCompound E3, wherein R₆ is hydrogen, lower alkyl, —(CH₂)_(n)—CF₃, loweralkoxy, alkoxycarbonyl, cyano or halogen.

To provide additional compounds of the present invention, Compound J3was used to replace Compound A6 and carried forward using the method ofScheme A to provide a target compound of Formula (I) andpharmaceutically acceptable anionic salt forms thereof.

Accordingly, using the procedure of Scheme A to provide additionalcompounds of the present invention, Compound E3 is used in place ofCompound A6 and carried forward to provide a target compound of Formula(I) and pharmaceutically acceptable anionic salt forms thereof.

The invention is further defined by reference to the following examples,which are intended to be illustrative and not limiting.

EXAMPLE 1[4-(3,4-dichloro-benzoylamino)-benzyl]-dimethyl-(tetrahydro-pyran-4-yl)-ammoniumchloride (Cpd 17)

After cooling a mixture of 4-nitrobenzylamine hydrochloride Compound 1a,(22.12 mmol, 4.18 g) and tetrahydro-4H-pyran-4-one Compound 1b (22.25mmol, 2.48 g) in CH₂Cl₂ (50 mL) to 0° C., Et₃N (22.43 mmol, 3.13 mL) wasadded, followed by NaB(OAc)₃H (31.0 mmol, 6.58 g). The resultingsuspension was allowed to warm to r.t. with stirring overnight. Analiquot of the reaction mixture showed the formation of product (MS m/e237, 100%). This portion of the procedure of Example 1 was adapted fromShiroshi, et al., J. Med. Chem., 2000, 43, 2049.

An aqueous solution of formaldehyde (37% solution, 24.32 mmol, 1.98 mL)was combined with Compound 1c, followed by NaB(OAc)₃H (31.0 mmol, 6.58g) added in one portion under ice cooling. The reaction mixture wasstirred at r.t. for 12 hrs, then made basic using a 2N NaOH solution andextracted with CH₂Cl₂. The organic layer was washed with brine, thenseparated and dried over Na₂SO₄. The drying agent was filtered and thesolvent was removed in vacuo to yieldmethyl-(4-nitro-benzyl)-(tetrahydro-pyran-4-yl)-amine Compound 1c as aorange-yellow thick oil (5.58 g), which was used in the next stepwithout further purification. This portion of the procedure of Example 1was adapted from Hashimoto et al., Org. Proc. R&D, 2002, 6, 70. MS m/e251 (M⁺H, 100%); ¹H NMR (300 MHz, CDCl₃) δ 1.60-1.82 (m, 4H), 2.21 (s,3H), 2.59-2.78 (m, 1H), 3.38 (dt, J=3.2 Hz, J=11.2 Hz, 2H), 3.68 (s,2H), 4.02-4.10 (m, 2H), 7.50 (d, J=8.8 Hz, 2H), 8.17 (d, J=8.8 Hz, 2H).

SnCl₂.2H₂O (71.2 mmol, 16.07 g) in conc. HCl (14 mL) was added in smallportions to a solution of Compound 1c (22.12 mmol, 5.58 g) in THF (10mL) at r.t. A mild exotherm was observed and the reaction mixture wasplaced in a water bath cooled with just enough ice to maintain r.t. Theresulting yellow solution was stirred for 1 hr. TLC analysis [9:1CH₂Cl₂:MeOH; R_(f) 0.6 (Compound 1c) and R_(f) 0.2 (Compound 1d)] showeda trace of starting material (Compound 1c). The round bottom flaskcontaining the reaction mixture was then placed in a Warm-water bath for30 min to allow the reaction to reach completion. The mixture wasdiluted with THF (50 mL) and water (30 mL) and made basic with 2N NaOHsolution. The layers were separated and the aqueous layer was extractedwith Et₂O (2×75 mL). The combined organic layers were dried over MgSO₄,filtered and the solvent was removed in vacuo to obtain(4-amino-benzyl)-methyl-(tetrahydro-pyran-4-yl)-amine Compound 1d as apale yellow solid. The solid was further dried by pressing it on afilter paper to obtain the product as an off-white powder (4.22 g, 86%yield over 3 steps). MS m/e 221 (M⁺H, 100%); ¹H NMR (400 MHz, CDCl₃) δ1.52-1.70 (m, 4H), 2.11 (s, 3H), 2.50-2.61 (m, 1H), 3.29 (dt, J=3.2 Hz,J=11.2 Hz, 2H), 3.42 (s, 2H), 3.54 (br, 2H), 3.91-3.98 (m, 2H), 6.58 (d,J=8.4 Hz, 2H), 7.10 (d, J=8.4 Hz, 2H).

A solution of 3,4-dichlorobenzoyl chloride Compound 1e (14.4 mmol, 3.01g) was added dropwise via a dropping funnel over 20 min to a solution ofCompound 1d (13.64 mmol, 3.0 g) and Et₃N (27.28 mmol, 3.8 mL) in THF(100 mL) at 0° C. The resulting suspension was allowed to warm to r.t.overnight, then made basic using a 2N NaOH solution and extracted withEtOAc (100 mL). The aqueous layer was extracted with EtOAc (2×50 mL).The organic layers were washed with brine, then dried over MgSO₄ andfiltered. The solvent was removed in vacuo to yield3,4-dichloro-N-(4-{[methyl-(tetrahydro-pyran-4-yl)-amino]-methyl}-phenyl)-benzamideCompound 1f as a yellow solid. The product was purified by flash columnchromatography (15:1 EtOAc:MeOH to 6:1 EtOAc:MeOH) to yield a whitepowder (4.6 g, 86%). Mp 135-136° C.; MS m/e 393 (M⁺H, 100%); ¹H NMR (300MHz, CDCl₃) δ 1.65-1.83 (m, 4H), 2.21 (s, 3H), 2.58-2.61 (m, 1H), 3.38(dt, J=3.1 Hz, J=11.0 Hz, 2H), 3.58 (s, 2H), 4.01-4.09 (m, 2H), 7.35 (d,J=8.4 Hz, 2H), 7.53-7.60 (m, 3H), 7.68-7.74 (m, 2H), 7.98 (d, J=3.0 Hz,1H); Anal. Calcd. for C₂₀H₂₂Cl₂N₂O₂: C, 61.08; H, 5.64; N, 7.12; Cl,18.03; Found C 61.09; H, 5.57; N, 6.93; Cl, 17.91.

Iodomethane (29.9 mmol, 18.6 mL) was added to a solution of Compound 1f(10.2 mmol, 4.0 g) in acetone (250 mL) and acetonitrile (100 mL) at r.t.The resulting solution was stirred overnight, after which a whiteprecipitate was observed. The solvent was removed in vacuo and theoff-white solid was washed with EtOAc (20 mL) and Et₂O (100 mL) thendried in a vacuum oven for 12 hrs to obtain[4-(3,4-dichloro-benzoylamino)-benzyl]-dimethyl-(tetrahydro-pyran-4-yl)-ammonium iodide Compound 1 g as a white solid (5.0 g, 91.7%). Mp210-213° C.; MS m/e 407 (M, 100%); ¹H NMR (300 MHz, CD₃OD) δ 1.92-2.10(m, 2H), 2.19-2.28 (m, 2H), 2.98 (s, 6H), 3.48 (t, J=11.4 Hz, 2H),3.65-3.77 (m, 1H), 4.10-4.19 (m, 2H), 4.54 (s, 2H), 7.58 (d, J=8.4 Hz,2H), 7.70 (d, J=8.4 Hz, 1H), 7.86-7.94 (m, 3H), 8.12 (d, J=2.1 Hz, 1H);Anal. Calcd. for C₂₁H₂₅Cl₂₁N₂O₂: C, 47.12; H, 4.71; N, 5.23, 123.71;Found C, 46.83; H, 4.57; N, 5.18, 123.38.

A solution of Compound 1 g (5.0 g) in 1:1 mixture of MeOH/H₂O was passedthrough a column packed with ˜300 g of anion-exchange resin (Bio-Rad,analytical grade anion exchange resin. AG 1-X8, 50-100 mesh, chlorideform). The column was washed with MeOH and some acetone and Et₂O wereadded to the flask. The solvent was removed in vacuo and the product wasdried in a vacuum oven overnight to provide Compound 17 as a whitepowder (3.9 g, 95%). Mp 227-232° C.; MS m/e 407 (M, 100%); ¹H NMR (400MHz, CD₃OD) δ 1.95-2.01 (m, 2H), 2.22-2.25 (m, 2H), 2.97 (s, 6H), 3.48(t, J=11.7 Hz, 2H), 3.65-3.72 (m, 1H), 4.14-4.18 (m, 2H), 4.53 (s, 2H),7.58 (d, J=8.6 Hz, 2H), 7.70 (d, J=8.4 Hz, 1H), 7.86-7.92 (m, 3H), 8.12(d, J=2.1 Hz, 1H); Anal. Calcd. for C₂₁H₂₅Cl₃N₂O₂: C, 56.83; H, 5.68; N,6.31; Cl, 23.97; Found C, 56.93; H, 5.72; N, 6.02; Cl, 23.67 (% I wasfound to be <0.1%).

Using the procedure of Example 1 and known appropriate reagents andstarting materials, other compounds of the present invention may beprepared including, (MS: Mass Spec data as MS m/e M⁺H):

Cpd Name MS 6(4-benzoylamino-benzyl)-dimethyl-(tetrahydro-pyran-4-yl)-ammonium 429iodide 9[4-(2,3-dichloro-benzoylamino)-benzyl]-dimethyl-(tetrahydro-pyran-4-yl)-407 ammonium iodide 10[4-(2,4-dichloro-benzoylamino)-benzyl]-dimethyl-(tetrahydro-pyran-4-yl)-407 ammonium iodide 11[4-(2,5-dichloro-benzoylamino)-benzyl]-dimethyl-(tetrahydro-pyran-4-yl)-407 ammonium iodide 12[4-(2,6-dichloro-benzoylamino)-benzyl]-dimethyl-(tetrahydro-pyran-4-yl)-407 ammonium iodide 13[4-(2-chloro-benzoylamino)-benzyl]-dimethyl-(tetrahydro-pyran-4-yl)- 373ammonium iodide 14bicyclo[2.2.1]hept-2-yl-[4-(3,4-dichloro-benzoylamino)-benzyl]-dimethyl-417 ammonium iodide 19[4-(3,4-dichloro-benzoylamino)-benzyl]-dimethyl-(tetrahydro-thien-3-yl)-409 ammonium iodide 20[4-(3,4-dichloro-benzoylamino)-benzyl]-dimethyl-(tetrahydro-thiopyran-4-423 yl)-ammonium iodide 21[4-(3,4-dichloro-benzoylamino)-benzyl]-ethyl-methyl-(tetrahydro-pyran-4-421 yl)-ammonium iodide 22[4-(3,4-dichloro-benzoylamino)-benzyl]-methyl-propyl-(tetrahydro-pyran-4-435 yl)-ammonium iodide 23[4-(3,5-dichloro-benzoylamino)-benzyl]-dimethyl-(tetrahydro-pyran-4-yl)-407 ammonium iodide 24[4-(3-bromo-benzoylamino)-benzyl]-dimethyl-(tetrahydro-pyran-4-yl)- 417ammonium iodide 25[4-(3-chloro-2-methyl-benzoylamino)-benzyl]-dimethyl-(tetrahydro-pyran-4-387 yl)-ammonium iodide 26[4-(3-chloro-4-fluoro-benzoylamino)-benzyl]-dimethyl-(tetrahydro-pyran-4-391 yl)-ammonium iodide 27[4-(3-chloro-4-methoxy-benzoylamino)-benzyl]-dimethyl-(tetrahydro-pyran-403 4-yl)-ammonium iodide 28[4-(3-chloro-4-methyl-benzoylamino)-benzyl]-dimethyl-(tetrahydro-pyran-4-387 yl)-ammonium iodide 29[4-(3-chloro-benzoylamino)-benzyl]-dimethyl-(tetrahydro-pyran-4-yl)- 359ammonium iodide 30[4-(3-cyano-benzoylamino)-benzyl]-dimethyl-(tetrahydro-pyran-4-yl)- 364ammonium iodide 31[4-(3-methoxy-benzoylamino)-benzyl]-dimethyl-(tetrahydro-pyran-4-yl)-369 ammonium iodide 32[4-(4-chloro-2-methyl-benzoylamino)-benzyl]-dimethyl-(tetrahydro-pyran-4-387 yl)-ammonium iodide 34[4-(4-chloro-benzoylamino)-benzyl]-dimethyl-(tetrahydro-pyran-4-yl)- 373ammonium iodide 35[4-(5-chloro-2-methyl-benzoylamino)-benzyl]-dimethyl-(tetrahydro-pyran-4-387 yl)-ammonium iodide 40{4-[3-(3,4-dichloro-phenyl)-acryloylamino]-benzyl}-dimethyl-(tetrahydro-419 pyran-4-yl)-ammonium iodide 41{4-[3-(3,4-dichloro-phenyl)-acryloylamino]-benzyl}-dimethyl-(tetrahydro-449 thiopyran-4-yl)-ammonium iodide 42{4-[3-(3,5-difluoro-phenyl)-acryloylamino]-benzyl}-dimethyl-(tetrahydro-401 pyran-4-yl)-ammonium iodide 43{4-[3-(3-bromo-phenyl)-acryloylamino]-benzyl}-dimethyl-(tetrahydro- 443pyran-4-yl)-ammonium iodide 44{4-[3-(3-bromo-phenyl)-acryloylamino]-benzyl}-dimethyl-(tetrahydro- 459thiopyran-4-yl)-ammonium iodide 45{4-[3-(3-chloro-phenyl)-acryloylamino]-benzyl}-dimethyl-(tetrahydro- 399pyran-4-yl)-ammonium iodide 46{4-[3-(3-fluoro-phenyl)-acryloylamino]-benzyl}-dimethyl-(tetrahydro- 383pyran-4-yl)-ammonium iodide 47{4-[3-(4-bromo-phenyl)-acryloylamino]-benzyl}-dimethyl-(tetrahydro- 443pyran-4-yl)-ammonium iodide 501-[4-(3,4-dichloro-benzoylamino)-benzyl]-1-methyl-piperidinium iodide377 51 1-[4-(3,4-dichloro-benzoylamino)-benzyl]-1-methyl-pyrrolidiniumiodide 363 541-{4-[3-(3,4-dichloro-phenyl)-acryloylamino]-benzyl}-4-(2-methoxy- 510phenyl)-1-methyl-piperazin-1-ium iodide 551-{4-[3-(3-bromo-phenyl)-acryloylamino]-benzyl}-1-methyl-piperidinium413 iodide 571-methyl-1-{4-[3-(3-trifluoromethyl-phenyl)-acryloylamino]-benzyl}- 403piperidinium iodide 594-[4-(3,4-dichloro-benzoylamino)-benzyl]-4-methyl-morpholin-4-ium iodide379 604-{4-[3-(3,4-dichloro-phenyl)-acryloylamino]-benzyl}-4-methyl-morpholin-405 4-ium iodide 614-{4-[3-(3-bromo-phenyl)-acryloylamino]-benzyl}-4-methyl-morpholin-4-415 ium iodide 63allyl-{4-[3-(3-bromo-phenyl)-acryloylamino]-benzyl}-methyl-(tetrahydro-487 thiopyran-4-yl)-ammonium iodide 66dimethyl-(tetrahydro-pyran-4-yl)-[4-(3-m-tolyl-acryloylamino)-benzyl]-379 ammonium iodide 67dimethyl-(tetrahydro-pyran-4-yl)-[4-(3-trifluoromethyl-benzoylamino)-407 benzyl]-ammonium iodide 68dimethyl-(tetrahydro-pyran-4-yl)-{4-[3-(3-trifluoromethyl-phenyl)- 433acryloylamino]-benzyl}-ammonium iodide 69dimethyl-[4-(3-methyl-benzoylamino)-benzyl]-(tetrahydro-pyran-4-yl)- 353ammonium iodide 70cycloheptyl-[4-(3,4-dichloro-benzoylamino)-benzyl]-dimethyl-ammonium 419iodide 71cyclohexyl-{4-[3-(3,4-dichloro-phenyl)-acryloylamino]-benzyl}-dimethyl-431 ammonium iodide 72{4-[3-(3-bromo-phenyl)-acryloylamino]-benzyl}-cyclohexyl-dimethyl- 443ammonium iodide 74cyclohexyl-dimethyl-[4-(3-trifluoromethyl-benzoylamino)-benzyl]- 405ammonium iodide 75cyclohexyl-[4-(3,4-dichloro-benzoylamino)-benzyl]-dimethyl-ammonium 405iodide 76[4-(3-chloro-4-fluoro-benzoylamino)-benzyl]-cyclohexyl-dimethyl- 389ammonium iodide 77cyclohexyl-[4-(2,3-dichloro-benzoylamino)-benzyl]-dimethyl-ammonium 405iodide 78cyclohexyl-[4-(2,6-dichloro-benzoylamino)-benzyl]-dimethyl-ammonium 405iodide 79[4-(3-chloro-4-methoxy-benzoylamino)-benzyl]-cyclohexyl-dimethyl- 401ammonium iodide 80[4-(3-chloro-4-methyl-benzoylamino)-benzyl]-cyclohexyl-dimethyl- 385ammonium iodide 81cyclohexyl-[4-(2,5-dichloro-benzoylamino)-benzyl]-dimethyl-ammonium 405iodide 82cyclopentyl-[4-(3,4-dichloro-benzoylamino)-benzyl]-dimethyl-ammonium 391iodide 89dimethyl-{4-[(naphthalene-1-carbonyl)-amino]-benzyl}-(tetrahydro-pyran-4-389 yl)-ammonium iodide 90dimethyl-{4-[(naphthalene-2-carbonyl)-amino]-benzyl}-(tetrahydro-pyran-4-389 yl)-ammonium iodide 91ethyl-methyl-{4-[(naphthalene-2-carbonyl)-amino]-benzyl}-(tetrahydro-403 pyran-4-yl)-ammonium iodide 92methyl-{4-[(naphthalene-2-carbonyl)-amino]-benzyl}-propyl-(tetrahydro-417 pyran-4-yl)-ammonium iodide 93{4-[(7-bromo-naphthalene-2-carbonyl)-amino]-benzyl}-dimethyl- 467(tetrahydro-pyran-4-yl)-ammonium iodide 94{4-[(7-bromo-naphthalene-2-carbonyl)-amino]-benzyl}-cyclohexyl- 465dimethyl-ammonium iodide 151dimethyl-{4-[(2-methyl-5-phenyl-furan-3-carbonyl)-amino]-benzyl}- 419(tetrahydro-pyran-4-yl)-ammonium iodide 152{4-[(benzofuran-2-carbonyl)-amino]-benzyl}-dimethyl-(tetrahydro-pyran-4-379 yl)-ammonium iodide 155{4-[(5-chloro-benzofuran-2-carbonyl)-amino]-benzyl}-dimethyl-(tetrahydro-413 pyran-4-yl)-ammonium iodide 156{4-[(5-chloro-benzofuran-2-carbonyl)-amino]-benzyl}-cyclohexyl-dimethyl-411 ammonium iodide 157{4-[(benzofuran-2-carbonyl)-amino]-benzyl}-cyclohexyl-dimethyl- 437ammonium iodide 172{4-[(5-bromo-pyridine-3-carbonyl)-amino]-benzyl}-dimethyl-(tetrahydro-420 pyran-4-yl)-ammonium iodide 173{4-[(2-chloro-pyridine-4-carbonyl)-amino]-benzyl}-dimethyl-(tetrahydro-374 pyran-4-yl)-ammonium iodide

EXAMPLE 2{2-[4-(3,4-dichloro-benzoylamino)-phenyl]-ethyl}-dimethyl-(tetrahydro-pyran-4-yl)-ammonium iodide (Cpd 36)

NaB(OAc)₃H (2.33 mmol, 0.5 g) was added to a mixture of2-(4-nitro-phenyl)-ethylamine Compound 2a (2.25 mmol, 0.45 g) andtetrahydro-4H-pyran-4-one Compound 1b (2.25 mmol, 0.21 mL) and Et₃N(2.25 mmol, 0.31 mL) in CH₂Cl₂ (25 mL). The resulting suspension wasstirred at r.t. for 12 hrs. An aliquot of the reaction mixture showedthe formation of product (MS m/e 251, 100%).

An aqueous solution of formaldehyde (37% solution, 8.6 mmol, 0.7 mL) wasadded, followed by NaB(OAc)₃H (2.33 mmol, 0.5 g) and the reactionmixture was stirred at r.t. for 12 hrs. The mixture was made basic usinga 2N NaOH solution, extracted with CH₂Cl₂ and the organic layer waswashed with brine, then separated and dried over Na₂SO₄. The dryingagent was filtered and the solvent was removed in vacuo to yieldmethyl-[2-(4-nitro-phenyl)-ethyl]-(tetrahydro-pyran-4-yl)-amine Compound2b as a yellow oil. The product was purified by flash columnchromatography (10:1 CH₂Cl₂:MeOH; R_(f) 0.8) to yield a yellow oil (0.58g, 97%). MS m/e 265 (M⁺H, 100%).

SnCl₂.2H₂O (10.0 mmol, 2.25 g) was added to a solution of Compound 2b(2.19 mmol, 0.58 g) in EtOH (10 mL) at r.t. A mild exotherm wasobserved. The resulting yellow solution was stirred for 12 hrs and thesolvent was removed in vacuo. The residue was made basic using a 2N NaOHsolution and the aqueous layer was extracted with CH₂Cl₂ (2×25 mL). Thecombined organic layers were dried over Na₂SO₄ and filtered, then thesolvent was removed in vacuo to obtain[2-(4-amino-phenyl)-ethyl]-methyl-(tetrahydro-pyran-4-yl)-amine Compound2c as an orange-yellow oil (0.4 g) used in the next step withoutpurification. MS m/e 235 (M⁺H, 100%).

A solution of 3,4-dichlorobenzoyl chloride Compound 1e (0.25 mmol, 0.06g) in THF (1 mL) was added dropwise over 2 min to a solution of Compound2c (0.2 mmol, 0.05 g) and Et₃N (0.4 mmol, 0.06 mL) in THF (4 mL) at 0°C. The resulting suspension was allowed to warm to r.t. overnight, thenmade basic with a 2N NaOH-solution and extracted with EtOAc (10 mL). Theaqueous layer was extracted with EtOAc (2×10 mL). The organic layerswere washed with brine, then dried over Na₂SO₄ and filtered. The solventwas removed in vacuo to yield3,4-dichloro-N-(4-{2-[methyl-(tetrahydro-pyran-4-yl)-amino]-ethyl)}-phenyl)-benzamideCompound 2d as a yellow solid. The product was purified by preparativeTLC (10:1 EtOAc:MeOH) (0.06 g, 73%). MS m/e 407 (M⁺H, 100%);

Iodomethane (0.5 mL, excess) was added to a solution of Compound 2d(0.07 mmol, 0.03 g) in acetone (1.0 mL) and acetonitrile (1.0 mL) atr.t. The resulting solution was stirred overnight, after which a yellowprecipitate was observed. The solvent was removed in vacuo and theoff-white solid was washed with Et₂O (2×5 mL) to provide Compound 36 asa pale yellow solid (0.03 g, 82%). MS m/e 548 (M, 100%).

Using the procedure of Example 2 and known appropriate reagents andstarting materials, other compounds of the present invention may beprepared including, (MS: Mass Spec data as MS m/e M⁺H):

Cpd Name MS 37 {2-[4-(3-bromo-benzoylamino)-phenyl]-ethyl}- 431dimethyl-(tetrahydro-pyran-4-yl)-ammonium iodide

EXAMPLE 3dimethyl-(tetrahydro-pyran-4-yl)-(4-{[3-(3-trifluoromethyl-phenyl)-acryloylamino]-methyl}-benzyl)-ammoniumiodide (Cpd 64)

Boc₂O was added in one portion at r.t. to a solution of(4-aminomethyl-phenyl)-methanol Compound 3a (21.2 mmol, 2.9 g) in CH₂Cl₂(100 mL). The resulting solution was stirred for 48 h, then washed witha 10% citric acid solution (50 mL) followed by brine. The organic-layerwas separated, then dried over Na₂SO₄ and filtered. The solvent wasremoved in vacuo to obtain (4-hydroxymethyl-benzyl)-carbamic acidtert-butyl ester Compound 3b as a white solid (5.2 g, 99% yield), whichwas used in the next step without further purification.

MnO₂ (9.6 g) was added to a solution of Compound 3b (21.2 mmol, 5.2 g)in chloroform (60 mL), forming a black suspension that was stirred atr.t. overnight then filtered through a pad of celite. The solvent wasevaporated in vacuo to obtain (4-formyl-benzyl)-carbamic acid tert-butylester Compound 3c as a white solid (4.3 g, 87% yield), which was used inthe next step without purification.

NaB(OAc)₃H (2.8 mmol, 0.58 g) was added to a mixture of Compound 3c (2.6mmol, 0.6 g) and tetrahydro-pyran-4-ylamine Compound 3d (2.6 mmol, 0.26g) in CH₂Cl₂ (25 mL) and the resulting suspension was stirred at r.t. Analiquot of the reaction mixture showed the formation of product (MS m/e321; 100%). An aqueous solution of formaldehyde (37% solution, 8.6 mmol,0.7 mL) was added to the reaction mixture, followed by NaB(OAc)₃H (2.8mmol, 0.58 g) added in one portion under ice cooling. The reactionmixture was stirred at r.t. for about 2 h, then made basic with a 2NNaOH solution and extracted with CH₂Cl₂. The organic layer was washedwith brine, then separated and dried over Na₂SO₄. The drying agent wasfiltered and the solvent was removed in vacuo to yield(4-{[methyl-(tetrahydro-pyran-4-yl)-amino]-methyl}-benzyl)-carbamic acidtert-butyl ester Compound 3e as a pale yellow oil. MS m/e 235 (M⁺H,100%). The product was purified by column chromatography (4:1CH₂Cl₂:MeOH) to yield a colorless oil (0.52 g, 59% yield).

Compound 3e was dissolved in CH₂Cl₂, then HCl in dioxane was added andthe mixture was stirred at r.t. for 12 hrs. The solvent was removed andthe gummy residue was made basic with 2N NaOH and extracted with EtOAc.The organic layer was washed with brine, then separated and dried overNa₂SO₄. The drying agent was filtered and the solvent was removed invacuo to obtain(4-aminomethyl-benzyl)-methyl-(tetrahydro-pyran-4-yl)-amine Compound 3fas a pale yellow oil (0.3 g, 83% yield). MS m/e 235 (M⁺H, 100%).

A solution of 3-(3-trifluoromethyl-phenyl)-acryloyl chloride Compound 3g(0.3 mmol, 0.07 g) in THF (2 mL) was added dropwise to a solution ofCompound 3f (0.2 mmol, 0.05 g) and Et₃N (0.8 mmol, 0.14 mL) in THF (10mL) at 0° C. The resulting suspension was allowed to warm to r.t.overnight. The reaction mixture was made basic with a 2N NaOH solutionand extracted with EtOAc (25 mL). The aqueous layer was extracted withEtOAc (2×10 mL) and the organic layers were washed with brine, thendried over Na₂SO₄ and filtered. The solvent was removed in vacuo toyield a yellow solid (with methane) as the product. The crude productwas purified by preparative TLC (9:1 EtOAc:MeOH, Rf=0.2) to yieldN-(4-{[methyl-(tetrahydro-pyran-4-yl)-amino]-methyl}-benzyl)-3-(3-trifluoromethyl-phenyl)-acrylamideCompound 3h (0.06 g, 49% yield). MS m/e 433 (M⁺H, 100%).

MeI (0.08 mL, 1.28 mmol) was added dropwise to a solution of Compound 3h(0.07 mmol, 0.03 g) in a mixture of acetone:acetonitrile (2 mL). Theresulting solution was stirred at r.t. for 24 h to provide a residue.The residue was washed with ether (2×1 mL) and dried under a high vacuumto provide Compound 64 (0.04 g, 93% yield) as an iodide salt. MS m/e 584(M⁺H, 100%).

Using the procedure of Example 3 and the appropriate known reagents andstarting materials, other compounds of the invention may be preparedincluding, (MS: Mass Spec data as MS m/e M⁺H),

Cpd Name MS 1(4-{[3-(3-bromo-phenyl)-acryloylamino]-methyl}-benzyl)-cyclohexyl- 455dimethyl-ammonium iodide 2{4-[(3-bromo-benzoylamino)-methyl]-benzyl}-cyclohexyl-dimethyl- 429ammonium iodide 3cyclohexyl-dimethyl-{4-[(3-trifluoromethyl-benzoylamino)-methyl]- 419benzyl}-ammonium iodide 4(4-{[3-(3,4-dichloro-phenyl)-acryloylamino]-methyl}-benzyl)-dimethyl-447 (tetrahydro-pyran-4-yl)-ammonium iodide 5(4-{[3-(3-bromo-phenyl)-acryloylamino]-methyl}-benzyl)-dimethyl- 457(tetrahydro-pyran-4-yl)-ammonium iodide 39{4-[(3,4-dichloro-benzoylamino)-methyl]-benzyl}-dimethyl-(tetrahydro-421 pyran-4-yl)-ammonium iodide 481-(4-{[3-(3,4-dichloro-phenyl)-acryloylamino]-methyl}-benzyl)-1-methyl-417 piperidinium iodide 491-(4-{[3-(3-bromo-phenyl)-acryloylamino]-methyl}-benzyl)-1-methyl- 427piperidinium iodide 584-(4-{[3-(3,4-dichloro-phenyl)-acryloylamino]-methyl}-benzyl)-4-methyl-419 morpholin-4-ium iodide 149dimethyl-(4-{[(2-methyl-5-phenyl-furan-3-carbonyl)-amino]-methyl}- 433benzyl)-(tetrahydro-pyran-4-yl)-ammonium iodide 150[4-({[5-(4-chloro-phenyl)-2-methyl-furan-3-carbonyl]-amino}-methyl)- 467benzyl]-dimethyl-(tetrahydro-pyran-4-yl)-ammonium iodide 153(4-{[5-(4-chloro-phenyl)-2-trifluoromethyl-furan-3-carbonyl]-amino}- 507benzyl)-dimethyl-(tetrahydro-pyran-4-yl)-ammonium iodide 154(4-{[5-(4-chloro-phenyl)-2-trifluoromethyl-furan-3-carbonyl]-amino}- 506benzyl)-cyclohexyl-dimethyl-ammonium iodide

EXAMPLE 4[4-(3,4-dichloro-benzoylamino)-benzyl]-dimethyl-(tetrahydro-pyran-4-ylmethyl)-ammoniumiodide (Cpd 18)

NaB(OAc)₃H (3.5 mmol, 0.75 g) was added to a mixture of4-nitro-benzaldehyde Compound 4a (2.8 mmol, 0.42 g),(tetrahydro-pyran-4-yl)-methylamine Compound 4b (3.0 mmol, 0.35 g) andglacial acetic acid (3 drops) in CH₂Cl₂ (50 mL) and the resultingsuspension was stirred at r.t. overnight. An aliquot of the reactionmixture showed the formation of product (MS m/e 251; 100%). An aqueoussolution of formaldehyde (37% solution, 9.6 mmol, 0.8 mL) was added,followed by NaB(OAc)₃H (3.5 mmol, 0.75 g). The reaction mixture wasstirred at r.t. for 2 h, then made basic with a 2N NaOH solution andextracted with CH₂Cl₂. The organic layer was washed with brine, thenseparated and dried over Na₂SO₄. The drying agent was filtered and thesolvent was removed in vacuo to yieldmethyl-(4-nitro-benzyl)-(tetrahydro-pyran-4-ylmethyl)-amine Compound 4cas yellow oil (0.63 g, 85% yield). MS m/e 265 (M⁺H, 100%).

SnCl₂.2H₂O (1.78 mmol, 0.4 g) was added to a solution of Compound 4c(1.13 mmol, 0.3 g) in EtOH (20 mL) at r.t. The resulting yellow solutionwas stirred for 2 days then the solvent was removed in vacuo. Theresidue was made basic with a 2N NaOH solution and the aqueous layer wasextracted with CH₂Cl₂ (2×25 mL). The combined organic layers were driedover Na₂SO₄ and filtered, then the solvent was removed in vacuo toobtain 4-{[methyl-(tetrahydro-pyran-4-ylmethyl)-amino]-methyl}-phenylamine Compound 4d as an orange-yellowoil (0.25 g, 94%) used in the next step without purification. MS m/e 235(M⁺H, 100%).

A solution of 3,4-dichlorobenzoyl chloride Compound 1e (0.29 mmol, 0.06g) in THF (1.0 mL) was added dropwise via syringe to a solution ofCompound 4d (0.19 mmol, 0.04 g) and Et₃N (0.36 mmol, 0.05 mL) in THF (4mL) at 0° C. The resulting suspension was allowed to warm to r.t.overnight, then made basic with a 2N NaOH solution and extracted withEtOAc (15 mL). The aqueous layer was extracted with EtOAc (2×10 mL). Theorganic layers were washed with brine, dried over Na₂SO₄ and thenfiltered. The solvent was removed in vacuo to yield3,4-dichloro-N-(4-{[methyl-(tetrahydro-pyran-4-ylmethyl)-amino]-methyl}-phenyl)-benzamideCompound 4e as a yellow solid. The product was purified by preparativeTLC (10:1 EtOAc:MeOH; Rf 0.4) (0.04 g, 52%). MS m/e 407 (M⁺H, 100%).

Iodomethane (0.5 mL) was added to a solution of Compound 4e (0.07 mmol,0.03 g) in acetone (1.0 mL) and acetonitrile (1.0 mL) at r.t. Theresulting solution was allowed to stand overnight, after which a yellowprecipitate was observed. The solvent was removed in vacuo and theyellow solid was washed with Et₂O (2×1 mL) to obtain Compound 18 as ayellow solid (0.03 g, 96%). MS m/e 421 (M, 100%).

EXAMPLE 51-{4-[3-(3,4-dichloro-phenyl)-acryloylamino]-benzyl}-1-methyl-piperidiniumiodide (Cpd 53)

NaB(OAc)₃H (6.6 mmol, 1.4 g) was added to a mixture of4-nitro-benzaldehyde Compound 4a (6.0 mmol, 0.9 g), piperidine Compound5a (9.0 mmol, 0.9 mL) and glacial acetic acid (5 drops) in CH₂Cl₂ (50mL) and the resulting suspension was stirred at r.t. overnight. Thereaction mixture was made basic with a 2N NaOH solution and extractedwith CH₂Cl₂. The organic layer was washed with brine, then separated anddried over Na₂SO₄. The drying agent was filtered and the solvent wasremoved in vacuo. The product was purified by flash columnchromatography (10:1 EtOAc:MeOH) to yield 1-(4-nitro-benzyl)-piperidineCompound 5b as a yellow oil (0.89 g, 67% yield). MS m/e 221 (M⁺H, 100%).

SnCl₂.2H₂O (10.0 mmol, 2.25 g) was added to a solution of Compound 5b(5.0 mmol, 1.1 g) in EtOH (25 mL) at r.t. and a mild exotherm wasobserved. The resulting yellow solution was stirred for 2 days then thesolvent was removed in vacuo. The residue was made basic with a 2N NaOHsolution and the aqueous layer was extracted with CH₂Cl₂ (2×25 mL). Thecombined organic layers were dried over Na₂SO₄ then filtered and thesolvent was removed in vacuo to provide4-piperidin-1-ylmethyl-phenylamine Compound 5c as an orange-yellow oil(0.8 g, 84% yield), which was used in the next step without furtherpurification. MS m/e 191 (M⁺H, 100%).

A solution of 3-(3,4-dichloro-phenyl)-acryloyl chloride Compound 5d(0.21 mmol, 0.05 g) in THF (1.0 mL) was added dropwise via syringe to asolution of Compound 5c (0.21 mmol, 0.04 g) and Et₃N (5.1 mmol, 0.7 mL)in THF (4 mL) at 0° C. The resulting suspension was allowed to warm tor.t. overnight, then made basic with a 2N NaOH solution and extractedwith EtOAc (25 mL). The aqueous layer was extracted with EtOAc (2×10mL). The organic layers were washed with brine, dried over Na₂SO₄ andthen filtered. The solvent was removed in vacuo to yield3-(3,4-dichloro-phenyl)-N-(4-piperidin-1-ylmethyl-phenyl)-acrylamideCompound 5e as a yellow solid. The product was purified by preparativeTLC (10:1 EtOAc:MeOH; Rf 0.4) to yield a yellow oil which was convertedto the hydrochloride salt by dissolving a solution of Compound 5e inCH₂Cl₂ with a solution of HCl in Et₂O, followed by removal of thesolvent (0.05 g, 60%). MS m/e 389 (M⁺H, 100%).

Iodomethane (0.5 mL) was added to a solution of Compound 5e (0.06 mmol,0.025 g) in acetone (1.0 mL) and acetonitrile (1.0 mL) at r.t. Theresulting solution was allowed to stand overnight, after which a yellowprecipitate was observed. The solvent was removed in vacuo and theyellow solid was washed with Et₂O (2×1 mL) to provide Compound 53 as ayellow solid (0.03 g, 89%). MS m/e 530 (M, 100%).

EXAMPLE 6[3-(3,4-dichloro-benzoylamino)-benzyl]-dimethyl-(tetrahydro-pyran-4-yl)-ammoniumiodide (Cpd 7)

NaB(OAc)₃H (11.87 mmol, 2.52 g) was added to a mixture of3-nitrobenzaldehyde Compound 6a (9.89 mmol, 1.49 g) and4-amino-tetrahydro-pyran Compound 3d (9.89 mmol, 1.00 g) in CH₂Cl₂ (50mL) and the resulting suspension was stirred overnight at r.t. Thereaction mixture was made basic with a 2N NaOH solution and extractedwith CH₂Cl₂. The organic layer was washed with brine, then separated anddried over MgSO₄. The drying agent was filtered and the solvent wasremoved in vacuo. The product was purified by flash columnchromatography (9:1 CH₂Cl₂:MeOH) to yield(3-nitro-benzyl)-(tetrahydro-pyran-4-yl)-amine Compound 6b as a yellowoil (1.91 g, 82%). (This portion of Example 6 was adapted from Shiroshi,et al., J. Med. Chem., 2000, 43, 2049). MS m/e 237 (M⁺H, 100%); ¹H NMR(300 MHz, CDCl₃) δ 1.35-1.53 (m, 4H), 1.82-1.95 (d, 2H), 2.65-2.8 (m,1H), 3.38 (dt, J=3.2 Hz, J=11.2 Hz, 2H), 3.92-4.05 (m, 4H), 7.45-7.54(t, 1H), 7.65-7.72 (d, 1H), 8.07-8.15 (d, 1H), 8.22 (s, 1H).

An aqueous solution of formaldehyde (37% solution, 9.4 mmol, 0.70 mL)was added to a solution of Compound 6b (8.09 mmol, 1.91 g) in CH₂Cl₂,followed by NaB(OAc)₃H (9.70 mmol, 2.06 g) added in one portion. Thereaction mixture was stirred at r.t. for 12 hrs. An aliquot of thereaction mixture showed the formation of product (MS m/e 251, 100%). Thereaction mixture was made basic with a 2N NaOH solution and extractedwith CH₂Cl₂. The organic layer was washed with brine, then separated anddried over MgSO₄. The drying agent was filtered and the solvent wasremoved in vacuo to yield methyl-(3-nitro-benzyl)-(tetrahydro-pyran-4-yl)-amine Compound 6c as a yellow oil (1.87 g), which was used in thenext step without further purification. MS m/e 251 (M⁺H, 100%); ¹H NMR(300 MHz, CDCl₃) δ 1.60-1.82 (m, 4H), 2.21 (s, 3H), 2.60-2.75 (m, 1H),3.38 (dt, J=3.2 Hz, J=11.2 Hz, 2H), 3.68 (s, 2H), 4.02-4.10 (m, 2H),7.45-7.54 (t, 1H), 7.65-7.72 (d, 1H), 8.07-8.15 (d, 1H), 8.22 s, 1H).

SnCl₂.2H₂O (14.868 mmol, 3.35 g) was added to a solution of Compound 6c(3.72 mmol, 0.930 g) in absolute ethanol (30 mL) at r.t. The reactionmixture was stirred overnight at 40° C. An aliquot of the reactionmixture showed the formation of product (MS m/e 221, 100%). The solventwas removed in vacuo to obtain an orange solid, which was made basic topH 9 with a 1N NaOH solution. The product was extracted with EtOAc, thendried over MgSO₄ and filtered. The solvent was removed in vacuo toobtain (3-amino-benzyl)-methyl-(tetrahydro-pyran-4-yl)-amine Compound 6das a yellow oil (0.490 g). MS m/e 221 (M⁺H, 100%); ¹H NMR (300 MHz,CDCl₃) δ 1.58-1.80 (m, 4H), 2.22 (s, 3H), 2.57-2.68 (m, 1H), 3.36 (dt,J=3.2 Hz, J=11.2 Hz, 2H), 3.50 (s, 2H), 3.65 (br, 2H), 3.98-4.10 (d,2H), 6.55-6.62 (d, 1H), 6.70 (m, 2H), 7.05-7.12 (t, 1H).

A solution of 3,4-dichlorobenzoyl chloride Compound 1e (0.250 mmol,0.0523 g) was added dropwise to a solution of Compound 6d (0.227 mmol,0.0500 g) and Et₃N (0.250 mmol, 0.04 mL) in THF (10 mL) at 0° C. Theresulting suspension was allowed to warm to r.t. overnight. An aliquotof the reaction mixture showed the formation of product (MS m/e 393,100%). The reaction mixture was made basic with a 2N NaOH solution andextracted with EtOAc. The organic layers were washed with brine, driedover MgSO₄ and then filtered. The solvent was removed in vacuo to yield3,4-dichloro-N-(3-{[methyl-(tetrahydro-pyran-4-yl)-amino]-methyl}-phenyl)-benzamideCompound 6e. The product was purified by preparative TLC (9:1EtOAc:MeOH) to yield a yellow solid (0.0380 g, 43%). MS m/e 393 (M⁺H,100%); ¹H NMR (300 MHz, CDCl₃) δ 1.58-1.83 (m, 4H), 2.21 (s, 3H),2.61-2.75 (m, 1H), 3.35 (dt, J=3.1 Hz, J=11.0 Hz, 2H), 3.57 (s, 2H),4.01-4.09 (m, 2H), 7.10 (d, J=1H), 7.25-7.32 (t, 1H), 7.45-7.52 (d, 1H),7.55-7.65 (d, 2H), 7.65-7.72 (m, 1H), 7.95 (m, 1H), 8.25 (s, 1H).

Iodomethane (0.0161 mol, 1.0 mL) was added to a solution of Compound 6e(0.0674 mmol, 0.0265 g) in acetonitrile (3 mL) and acetone (3 drops) atr.t. The resulting solution was stirred overnight, then the solvent wasremoved in vacuo. The product was washed with Et₂O (10 mL) and dried ina vacuum oven for 12 hrs to provide Compound 7 as an orange solid(0.0326 g, 90.3%). MS m/e 407 (M, 100%).

Using the procedure of Example 6 and appropriate known reagents andstarting materials, other compounds of the present invention may beprepared including, but not limited to (MS: Mass Spec data as MS m/eM⁺H):

Cpd Name MS 8 [3-(3-bromo-benzoylamino)-benzyl]-dimethyl-(tetrahydro-419 pyran-4-yl)-ammonium iodide 38{3-[3-(3-bromo-phenyl)-acryloylamino]-benzyl}- 445dimethyl-(tetrahydro-pyran-4-yl)-ammonium iodide 521-{3-[3-(3-bromo-phenyl)-acryloylamino]-benzyl}- 4151-methyl-piperidinium iodide 561-methyl-1-[3-(3-trifluoromethyl-benzoylamino)-benzyl]- 377 piperidiniumiodide 65 dimethyl-(tetrahydro-pyran-4-yl)-[3-(3-trifluoromethyl- 407benzoylamino)-benzyl]-ammonium iodide 83cyclohexyl-{3-[3-(3,4-dichloro-phenyl)-acryloylamino]- 431benzyl}-dimethyl-ammonium iodide 84cyclohexyl-{3-[3-(4-fluoro-phenyl)-acryloylamino]- 381benzyl}-dimethyl-ammonium iodide

EXAMPLE 7[4-(4-chloro-3-trifluoromethyl-benzoylamino)-benzyl]-dimethyl-(tetrahydro-pyran-4-yl)-ammoniumiodide (Cpd 33)

EDIC hydrochloride (0.33 mmol, 0.07 g) was added in one portion to asuspension of (4-amino-benzyl)-methyl-(tetrahydro-pyran-4-yl)-amineCompound 1d (0.25 mmol, 0.06 g), 4-chloro-3-trifluoromethyl-benzoic acidCompound 7a (0.22 mmol, 0.05 g) and HOBt (0.22 mmol 0.03 g) in DMF (5.0mL) at 0° C. The resulting suspension was warmed to r.t. and a crystalof DMAP and Et₃N (0.65 mmol, 0.1 mL) were added. The mixture was stirredovernight and produced an orange-yellow suspension. The suspension waspoured into water and the aqueous layer was extracted with EtOAc (25mL). The organic layer was washed with water (2×20 mL), then a solutionof 5% NaOH (10 mL) and brine. The organic layer was separated, driedover Na₂SO₄ and filtered. The solvent was removed in vacuo to yield aresidue, which was purified via preparative TLC (15:1 CH₂Cl₂:MeOH) toprovide4-chloro-N-(4-{[methyl-(tetrahydro-pyran-4-yl)-amino]-methyl}-phenyl)-3-trifluoromethyl-benzamideCompound 7b (0.06 g, 63%) as a pale yellow solid. MS m/e 427 (M⁺H,100%).

Iodomethane (0.5 mL) was added to a solution of Compound 7b (0.07 mmol,0.03 g) in CH₂Cl₂ (1.0 mL) at r.t. The resulting solution stoodovernight, after which a pale yellow precipitate was observed. Thesolvent was removed in vacuo and the yellow solid was washed with Et₂O(2×1 mL) to provide Compound 33 as a yellow solid (0.03 g, 96%). MS m/e441 (M⁺H, 100%).

EXAMPLE 8(2S)-[4-(3,4-dichloro-benzoylamino)-benzyl]-dimethyl-(tetrahydro-furan-2-ylmethyl)-ammonium iodide (Cpd 15)

NaB(OAc)₃H (3.5 mmol, 0.75 g) was added to a mixture of4-nitro-benzaldehyde Compound 4a (2.8 mmol, 0.42 g),(S)-(+)-(tetrahydro-furan-2-yl)-methylamine Compound 8a (3.0 mmol, 0.3mL) and glacial acetic acid (2 drops) in CH₂Cl₂ (25 mL) and theresulting suspension was allowed to stir at room temperature for 12 hrs.An aliquot of the reaction mixture showed the formation of(S)-(4-nitro-benzyl)-(tetrahydro-furan-2-ylmethyl)-amine Compound 8b (MSm/e 237, 100%).

An aqueous solution of formaldehyde (37% solution, 9.6 mmol, 0.8 mL) wasadded to the reaction mixture followed by NaB(OAc)₃H (3.5 mmol, 0.75 g)and the reaction mixture was allowed to stir at r.t. for 2 hrs. Thereaction mixture was basified with 2N NaOH solution and was extractedwith CH₂Cl₂. The organic layer was washed with brine, separated anddried over Na₂SO₄. The drying agent was filtered and the solvent wasremoved in vacuo. The gummy residue thus obtained was spectroscopicallycharacterized to be(S)-methyl-(4-nitro-benzyl)-(tetrahydro-furan-2-ylmethyl)-amine Compound8c (0.74 g). MS m/e 251 (M⁺H, 100%).

SnCl₂.2H₂O (10.0 mmol, 2.35 g) was added to a solution of Compound 8c(2.8 mmol, 0.74 g) in EtOH (25 mL) at r.t. and the resulting yellowsolution was stirred overnight. The solvent was removed in vacuo. Theresidue was basified with 2N NaOH solution and the aqueous layer wasextracted with CH₂Cl₂ (2×30 mL). The combined organic layers were driedover Na₂SO₄, filtered and the solvent was removed in vacuo to obtain(S)-4-{[methyl-(tetrahydro-furan -2-ylmethyl)-amino]-methyl}-phenylamineCompound 8d as a thick yellow oil (0.54 g, 88% yield), which was used inthe next step without purification. MS m/e 221 (M⁺H, 100%).

A solution of 3,4-dichlorobenzoyl chloride Compound 1e (0.25 mmol, 0.05g) in THF (5 mL) was added to a solution of Compound 8d (0.25 mmol, 0.06g) and Et₃N (0.5 mmol, 0.07 mL) in THF (3 mL) at 0° C. and the reactionmixture was stirred overnight. The pale yellow suspension was poured inwater and was extracted with EtOAc (20 mL). The organic layer was washedwith water (2×20 mL) followed by brine. The organic layer was separated,dried over Na₂SO₄ and filtered. The solvent was removed in vacuo and theresulting residue was purified by preparative TLC (15:1 CH₂Cl₂/MeOH) toyield (S)-3,4-dichloro-N-(4-{[methyl-(tetrahydro-furan-2-ylmethyl)-amino]-methyl}-phenyl)-benzamide Compound 8e as a paleyellow solid (0.06 g, 61%). M.S. m/e 393 (M⁺H, 100%).

Iodomethane (0.5 mL) was added to a solution of Compound 8e (0.07 mmol,0.03 g) in CH₂Cl₂ (1.0 mL) at r.t. and the resulting solution wasallowed to stand overnight. A yellow precipitate was observed. Thesolvent was removed in vacuo and the yellow solid was washed with Et₂Oto obtain Compound 15 as a off-white powder (0.04 g, 97%). MS m/e 534(M, 100%).

Using the procedure of Example 8 and the appropriate known reagents andstarting materials, other compounds of the invention may be preparedincluding, (MS: Mass Spec data as MS m/e M⁺H):

Cpd Name MS 16 (2R)-[4-(3,4-dichloro-benzoylamino)-benzyl]- 407dimethyl-(tetrahydro-furan-2-ylmethyl)-ammonium iodide 165(2S)-{4-[(5-chloro-1H-indole-2-carbonyl)- 412amino]-benzyl}-dimethyl-(tetrahydro-furan- 2-ylmethyl)-ammonium iodide

EXAMPLE 94-methyl-4-(4-{[3-(3-trifluoromethyl-phenyl)-acryloylamino]-methyl}-benzyl)-morpholin-4-iumiodide (Cpd 62)

NaB(OAc)₃H (0.82 mmol, 0.17 g) was added to a mixture of(4-formyl-benzyl)-carbamic acid tert-butyl ester Compound 3c (0.75 mmol,0.17 g) and morpholine Compound 9a (0.75 mmol, 0.07 mL) in CH₂Cl₂ (20mL) and the resulting suspension was stirred at room temperature for 6hrs. The reaction mixture was basified with 2N NaOH solution and wasextracted with CH₂Cl₂. The organic layer was washed with brine,separated and dried over Na₂SO₄. The drying agent was filtered and thesolvent was removed in vacuo to a crude product as a pale yellow oil. MSm/e 307 (M⁺H, 100%). The product was purified by prep TLC (10:1CH₂Cl₂/MeOH, Rf=0.5) to yield (4-morpholin-4-ylmethyl-benzyl)-carbamicacid tert-butyl ester Compound 9b.

Compound 9b was dissolved in CH₂Cl₂ and was stirred with HCl in dioxaneat r.t. for 12 hrs. The solvent was removed to obtain a gummy residue,which was basified with 2N NaOH and extracted with EtOAc. The organiclayer was washed with brine, separated and dried over Na₂SO₄. The dryingagent was filtered and the solvent was removed in vacuo to obtain4-morpholin-4-ylmethyl-benzylamine Compound 9c as pale yellow oil (wt.0.09 g, 58% yield). MS m/e 207 (M⁺H, 100%).

A solution of 3-(3-trifluoromethyl-phenyl)-acryloyl chloride Compound 3g(0.3 mmol, 0.07 g) in THF (2 mL) was added dropwise to a solution ofCompound 9c (0.19 mmol, 0.04 g) and Et₃N (0.8 mmol, 0.14 mL) in THF (10mL) at 0° C. The resulting suspension was allowed to warm to r.t.overnight. The reaction mixture was basified with 2N NaOH solution andwas extracted with EtOAc (25 mL). The aqueous layer was extracted withEtOAc (2×10 mL). The organic layers were washed with brine, then driedover Na₂SO₄ and filtered. The solvent was removed in vacuo and theresulting yellow gummy oil was purified by preparative TLC (10:1CH₂Cl₂/MeOH, Rf=0.5) to yieldN-(4-morpholin-4-ylmethyl-benzyl)-3-(3-trifluoromethyl-phenyl)-acrylamideCompound 9d as a pale yellow solid (0.06 g, 77%). MS m/e 405 (M⁺H,100%).

MeI (1.28 mmol, 0.08 mL) was added dropwise to a solution of Compound 9d(0.07 mmol, 0.03 g) in a mixture of acetone/acetonitrile (2 mL). Thesolution was stirred at room temperature for 24 hrs and concentrated.The resulting residue was washed with ether (2×1 mL) and dried underhigh vacuum to give Compound 62 (0.03 g, 78%). MS m/e 546 (M).

EXAMPLE 10[4-(3-bromo-benzoylamino)-benzyl]-cyclohexyl-dimethyl-ammonium iodide(Cpd 73)

NaB(OAc)₃H (11.0 mmol, 2.33 g) was added to a mixture of4-nitro-benzaldehyde Compound 4a (10.0 mmol, 1.51 g), cyclohexylamineCompound 10a (10.5 mmol, 1.2 mL) and glacial acetic acid (5 drops) inCH₂Cl₂ (40 mL) and the resulting suspension was allowed to stir at roomtemperature for 12 hrs. An aliquot of the reaction mixture showed theformation of product (MS m/e 235, 100%). The reaction mixture wasbasified with 2N NaOH solution and was extracted with CH₂Cl₂. Theorganic layer was washed with brine, separated and dried over Na₂SO₄.The drying agent was filtered and the solvent was removed in vacuo toyield cyclohexyl-(4-nitro-benzyl)-amine Compound 10b as yellow oil (1.56g, 67% yield), which was used in the next step without purification.

An aqueous solution of formaldehyde (37% solution, 9.6 mmol, 0.8 mL) wasadded to a solution of Compound 10b (3.41 mmol, 0.8 g) in CH₂Cl₂followed by NaB(OAc)₃H (7.0 mmol, 1.5 g) and the mixture was allowed tostir at r.t. for 2 hrs. The reaction mixture was basified with 2N NaOHsolution and was extracted with CH₂Cl₂. The organic layer was washedwith brine, separated and dried over Na₂SO₄. The drying agent wasfiltered and the solvent was removed in vacuo. The gummy residue waspurified by column chromatography (9:1 EtOAc/MeOH) to yieldcyclohexyl-methyl-(4-nitro-benzyl)-amine Compound 10c as yellow oil (0.8g, 94% yield). MS m/e 249 (M⁺H, 100%).

SnCl₂.2H₂O (16.0 mmol, 3.6 g) was added to a solution of Compound 10c(3.2 mmol, 0.8 g) in EtOH (40 mL) at r.t. and the resulting yellowsolution was stirred overnight. The solvent was removed in vacuo. Theresidue was basified with 2N NaOH solution and the aqueous layer wasextracted with CH₂Cl₂ (2×30 mL). The combined organic layers were driedover Na₂SO₄, filtered and the solvent was removed in vacuo to obtain4-[(cyclohexyl-methyl-amino)-methyl]-phenylamine Compound 10d as a thickyellow oil (0.69 g, 98% yield), which was used in the next step withoutpurification. MS m/e 219 (M⁺H, 100%).

A solution of 3-bromobenzoyl chloride Compound 10e (0.8 mmol, 0.15 g)was added to a solution of Compound 10d (0.7 mmol, 0.2 g) and Et₃N (0.8mmol, 0.14 mL) in THF (15 mL) in THF (5 mL) at 0° C. and the reactionmixture was stirred overnight. The pale yellow suspension was poured inwater and was extracted with EtOAc (30 mL). The organic layer was washedwith water (2×20 mL) followed by 5% NaOH solution (10 mL) and brine. Theorganic layer was separated, dried over Na₂SO₄ and filtered. The solventwas removed in vacuo and the resulting residue was purified bypreparative TLC (15:1 CH₂Cl₂/MeOH) to yield3-bromo-N-{4-[(cyclohexyl-methyl-amino)-methyl]-phenyl}-benzamideCompound 10f as a pale yellow solid (0.21 g, 75%). MS m/e 401 (M⁺H,100%).

Iodomethane (0.5 mL) was added to a solution of Compound 10f (0.07 mmol,0.03 g) in CH₂Cl₂ (1.0 mL) at r.t. and the resulting solution wasallowed to stand overnight. A yellow precipitate was observed. Thesolvent was removed in vacuo and the yellow solid was washed with Et₂Oto provide Compound 73 as a pale yellow solid (0.04 g, 99%). MS m/e 542(M, 100%).

EXAMPLE 11cyclohexyl-dimethyl-{4-[(4′-methyl-biphenyl-3-carbonyl)-amino]-benzyl}-ammoniumiodide (Cpd 86)

K₂CO₃ (0.2 mmol, 0.03 g) and a Pd(dppf)₂ catalyst:DCM complex (1:1)(0.03 mmol, 0.02 g) was added to a mixture of3-bromo-N-{4-[(cyclohexyl-methyl-amino)-methyl]-phenyl}-benzamideCompound 10f (0.1 mmol, 0.04 g) and p-tolylboronic acid Compound 11a(0.12 mmol, 0.02 g) in a mixed solution of toluene/ethanol/water (7 mL/1mL/1 mL). The resulting suspension was heated to reflux for 5 hrs,concentrated and purified with preparative TLC (10% MeOH/2% Et₃N/88%EtOAc) to yield 4′-methyl-biphenyl-3-carboxylic acid{4-[(cyclohexyl-methyl-amino)-methyl]-phenyl}amide Compound 11b (0.02 g,48%). MS m/e 413 (M+1).

Iodomethane (0.32 mmol, 0.02 mL) was added dropwise to a solution ofCompound 11b (0.012 mmol, 0.005 g) in acetone/acetonitrile (1 mL, 0.5mL/0.5 mL). The resulting solution was stirred at room temperature for48 hrs and concentrated. The obtained residue was washed with ether (2×1mL) and dried under a high vacuum to give Compound 86 (0.01 g, 89%). MSm/e 427 (M⁺H).

Using the procedure of Example 11 and known appropriate reagents andstarting materials, other compounds of the present invention may beprepared including, (MS: Mass Spec data as MS m/e M⁺H):

Cpd Name MS 85 dimethyl-(tetrahydro-pyran-4-yl)-{4-[(4′-trifluoromethyl-483 biphenyl-3-carbonyl)-amino]-benzyl}-ammonium iodide 87dimethyl-{4-[(4′-methyl-biphenyl-3-carbonyl)-amino]- 429benzyl}-(tetrahydro-pyran-4-yl)-ammonium iodide 88{4-[(biphenyl-4-carbonyl)-amino]-benzyl}- 415dimethyl-(tetrahydro-pyran-4-yl)-ammonium iodide

EXAMPLE 12dimethyl-{4-[(1-methyl-1H-indole-2-carbonyl)-amino]-benzyl}-(tetrahydro-pyran-4-yl)-ammoniumiodide (Cpd 158)

EDIC (0.33 mmol, 0.07 g) was added in one portion to a mixture of(4-amino-benzyl)-methyl-(tetrahydro-pyran -4-yl)-amine Compound 1d (0.25mmol, 0.06 g), 1-methyl-1H-indole-2-carboxylic acid Compound 12a (0.22mmol, 0.04 g) and HOBt (0.22 mmol, 0.03 g) in DMF (5.0 mL) at 0° C. Theresulting suspension was warmed to r.t. and then a crystal of DMAP andEt₃N (0.65 mmol, 0.11 mL) was added and the reaction mixture was stirredovernight. The resulting orange-yellow suspension was poured in waterand was extracted with EtOAc (25 mL). The organic layer was washed withwater (2×20 mL) followed by 5% NaOH solution (10 mL) and brine. Theorganic layer was separated, dried over Na₂SO₄ and filtered. The solventwas removed in vacuo and the resulting residue was purified bypreparative TLC (15:1 CH₂Cl₂/MeOH) to yield1-methyl-1H-indole-2-carboxylic acid (4-{[methyl-(tetrahydro-pyran-4-yl)-amino]-methyl}-phenyl)-amide Compound 12b as a pale yellow solid(0.05 g, 60%). MS m/e 378 (M⁺H, 100%).

Iodomethane (0.5 mL) was added to a solution of Compound 12b (0.08 mmol,0.03 g) in CH₂Cl₂ (1.0 mL) at r.t. The mixture was allowed to standovernight and a yellow precipitate was observed. The solvent was removedin vacuo and the yellow solid was washed with Et₂O to obtain Compound158 as a yellow solid (0.03 g, 77%). MS m/e 391 (M⁺H, 100%).

Using the procedure of Example 12 and known appropriate reagents andstarting materials, other compounds of the present invention may beprepared including, (MS: Mass Spec data as MS m/e M⁺H):

Cpd Name MS 159{4-[(5-chloro-1H-indole-2-carbonyl)-amino]-benzyl}-dimethyl-(tetrahydro-412 pyran-4-yl)-ammonium iodide 160{4-[(5-bromo-1H-indole-2-carbonyl)-amino]-benzyl}-dimethyl-(tetrahydro-456 pyran-4-yl)-ammonium iodide 161dimethyl-{4-[(1-methyl-1H-indole-3-carbonyl)-amino]-benzyl}-(tetrahydro-392 pyran-4-yl)-ammonium iodide 163cyclohexyl-dimethyl-{4-[(1-methyl-1H-indole-2-carbonyl)-amino]-benzyl}-390 ammonium iodide 166bicyclo[2.2.1]hept-2-ylmethyl-{4-[(5-chloro-1H-indole-2-carbonyl)-amino]-422 benzyl}-dimethyl-ammonium iodide 174{4-[(3-chloro-benzo[b]thiophene-2-carbonyl)-amino]-benzyl}-dimethyl- 429(tetrahydro-pyran-4-yl)-ammonium iodide 175{4-[(2,5-dichloro-thiophene-3-carbonyl)-amino]-benzyl}-dimethyl- 413(tetrahydro-pyran-4-yl)-ammonium iodide 176{4-[(benzo[b]thiophene-2-carbonyl)-amino]-benzyl}-dimethyl-(tetrahydro-395 pyran-4-yl)-ammonium iodide 177{4-[(benzo[b]thiophene-2-carbonyl)-amino]-benzyl}-cyclohexyl-dimethyl-393 ammonium iodide 178{4-[(3-chloro-benzo[b]thiophene-2-carbonyl)-amino]-benzyl}-cyclohexyl-427 dimethyl-ammonium iodide

EXAMPLE 13{4-[(1-benzyl-1H-indole-3-carbonyl)-amino]-benzyl}-dimethyl-(tetrahydro-pyran-4-yl)-ammoniumiodide (Cpd 162)

EDCI (0.33 mmol, 0.07 g) was added in one portion to a suspension of(4-amino-benzyl)-methyl-(tetrahydro-pyran -4-yl)-amine Compound 1d (0.25mmol, 0.06 g), 1-benzyl-1H-indole-3-carboxylic acid Compound 13a (0.22mmol, 0.06 g) and HOBt (0.22 mmol, 0.03 g) in DMF (5.0 mL) at 0° C. Theresulting suspension was warmed to r.t. and then a crystal of DMAP andEt₃N (0.65 mmol, 0.1 mL) was added and the reaction mixture was stirredovernight. The orange-yellow suspension was poured in water and wasextracted with EtOAc (25 mL). The organic layer was washed with water(2×20 mL) followed by 5% NaOH solution (10 mL) and brine. The organiclayer was separated, dried over Na₂SO₄ and filtered. The solvent wasremoved in vacuo and the resulting residue was purified by preparativeTLC (15:1 CH₂Cl₂/MeOH) to yield Compound 13b as a pale yellow solid(0.07 g, 71%). MS m/e 454 (M⁺H, 100%).

Iodomethane (0.5 mL) was added to a solution of Compound 13b (0.08 mmol,0.04 g) in CH₂Cl₂ (1.0 mL) at r.t. The mixture was allowed to standovernight and a yellow precipitate was observed. The solvent was removedin vacuo and the yellow solid was washed with Et₂O to obtain Compound162 as a yellow solid (0.05 g, 84%). MS m/e 469 (M⁺H, 100%).

EXAMPLE 14{4-[(5-chloro-1H-indole-2-carbonyl)-amino]-benzyl}-cyclohexyl-dimethyl-ammoniumiodide (Cpd 164)

Sodium triacetoxyborohydride (11.0 mmol, 2.33 g) was added to a mixtureof 4-nitro-benzaldehyde Compound 4a (10.0 mmol, 1.51 g), cyclohexylamineCompound 10a (10.5 mmol, 1.2 mL) and glacial acetic acid (5 drops) inCH₂Cl₂ (40 mL) and the resulting suspension was allowed to stir at roomtemperature for 12 hrs. An aliquot of the reaction mixture showed theformation of product (MS m/e 235, 100%). The reaction mixture wasbasified with 2N NaOH solution and was extracted with CH₂Cl₂. Theorganic layer was washed with brine, separated and dried over Na₂SO₄.The drying agent was filtered and the solvent was removed in vacuo toyield cyclohexyl-(4-nitro-benzyl)-amine Compound 14a (1.56 g, 67%) as ayellow oil, which was used in the next step without furtherpurification.

An aqueous solution of formaldehyde (37% solution, 9.6 mmol, 0.8 mL) wasadded to a solution of Compound 14a (3.41 mmol, 0.8 g) in CH₂Cl₂,followed by sodium triacetoxyborohydride (7.0 mmol, 1.5 g). The mixturewas allowed to stir at r.t. for 2 hrs. The reaction mixture was basifiedwith 2N NaOH solution and was extracted with CH₂Cl₂. The organic layerwas washed with brine, separated and dried over Na₂SO₄. The drying agentwas filtered and the solvent was removed in vacuo. The resulting gummyresidue was purified by column chromatography (9:1 EtOAc/MeOH) to yieldcyclohexyl-methyl-(4-nitro-benzyl)-amine Compound 14b (0.8 g, 94%) as ayellow oil. MS m/e 249 (M⁺H, 100%).

SnCl₂.2H₂O (16.0 mmol, 3.6 g) was added to a solution of Compound 14b(3.2 mmol, 0.8 g) in EtOH (40 mL) at r.t. The resulting yellow solutionwas stirred overnight and the solvent was removed in vacuo. Theresulting residue was basified with 2N NaOH solution and the aqueouslayer was extracted with CH₂Cl₂ (2×30 mL). The combined organic layerswere dried over Na₂SO₄, filtered and the solvent was removed in vacuo toobtain 4-[(cyclohexyl-methyl-amino)-methyl]-phenylamine Compound 14c(0.69 g, 98%) as a thick yellow oil, which was used in the next stepwithout further purification. MS m/e 219 (M⁺H, 100%).

EDCI (0.33 mmol, 0.07 g) was added in one portion to a suspension ofCompound 14c (0.25 mmol, 0.05 g), 5-chloro-1H-indole-2-carboxylic acidCompound 14d (0.22 mmol, 0.04 g) and HOBt (0.22 mmol, 0.03 g) in DMF(5.0 mL) at 0° C. The resulting suspension was warmed to r.t. and then acrystal of DMAP and Et₃N (0.65 mmol, 0.1 mL) was added and the reactionmixture was stirred overnight. The resulting orange-yellow suspensionwas poured in water and was extracted with EtOAc (25 mL). The organiclayer was washed with water (2×20 mL) followed by 5% NaOH solution (10mL) and brine. The organic layer was separated, dried over Na₂SO₄ andfiltered. The solvent was removed in vacuo and the resulting residue waspurified by preparative TLC (15:1 CH₂Cl₂/MeOH) to yield5-chloro-1H-indole-2-carboxylic acid{4-[(cyclohexyl-methyl-amino)-methyl]-phenyl}-amide Compound 14e (0.06g, 68%) as a pale yellow solid. MS m/e 396 (M⁺H, 100%).

Iodomethane (0.5 mL) was added to a solution of Compound 14e (0.08 mmol,0.03 g) in CH₂Cl₂ (1.0 mL) at r.t. The resulting solution was allowed tostand overnight and a yellow precipitate was observed. The solvent wasremoved in vacuo and the resulting yellow solid was washed with Et₂O toobtain Compound 167 as a yellow solid (0.04 g, 72%). MS m/e 410 (M⁺H,100%).

EXAMPLE 15(2S)-bicyclo[2.2.1]hept-2-yl-{4-[(6-chloro-2H-chromene-3-carbonyl)-amino]-benzyl}-dimethyl-ammoniumiodide (Cpd 110)

5-chloro-2-hydroxy-benzaldehyde Compound 15a (10.0 mmol, 1.7 g),acrylonitrile (50.0 mmol, 2.14 mL) and DABCO (2.33 mmol, 0.26 g) weremixed together and heated to reflux overnight using an oil bath. Afterthe flask was cooled to room temperature, Et₂O (100 mL) was added andthe Et₂O layer was washed with 10% NaOH solution followed by 1N HCl andbrine. The organic layer was dried over MgSO₄, filtered and the solventwas removed in vacuo to obtain 6-chloro-2H-chromene-3-carbonitrileCompound 15b as a yellow solid (1.42 g, 74%), which was used in the nextstep without further purification (the preceding was described in Wise,L. et al. J. Med. Chem., 1988, 31, 688).

THF (2 mL) and 10% NaOH solution (100 mL) was added to a round bottomflask containing Compound 15b (7.43 mmol, 1.42 g). The solution washeated to reflux for 4 hrs. The flask was immersed in an ice-bath andthe solution was acidified by careful addition of conc. HCl. Theresulting pale yellow solid was filtered and dried in a vacuum oven toobtain 6-chloro-2H-chromene-3-carboxylic acid Compound 15c (1.02 g,65%).

Sodium triacetoxyborohydride (3.5 mmol, 0.75 g) was added to a mixtureof 4-nitro-benzaldehyde Compound 4a (2.8 mmol, 0.42 g),(2S)-bicyclo[2.2.1]hept-2-ylamine Compound 15d (3.0 mmol, 0.33 g) andglacial acetic acid (3 drops) in CH₂Cl₂ (40 mL). The resultingsuspension was allowed to stir at room temperature for 12 hrs. Analiquot of the reaction mixture showed the formation of product (MS m/e247, 100%). An aqueous solution of formaldehyde (37% solution, 9.6 mmol,0.8 mL) was added to the reaction mixture followed by sodiumtriacetoxyborohydride (3.5 mmol, 0.75 g) and the mixture was allowed tostir at r.t. for 2 hrs. The reaction mixture was basified with 2N NaOHsolution and was extracted with CH₂Cl₂. The organic layer was washedwith brine, separated and dried over Na₂SO₄. The drying agent wasfiltered and the solvent was removed in vacuo to obtain(2S)-bicyclo[2.2.1]hept-2-yl-methyl-(4-nitro-benzyl)-amine Compound 15e(0.72 g, 98%) as an orange oil. MS m/e 261 (M⁺H, 100%), which was usedin the next step without further purification.

SnCl₂.2H₂O (10.4 mmol, 2.35 g) was added to a solution of Compound 15e(2.76 mmol, 0.72 g) in EtOH (25 mL) at r.t. The resulting yellowsolution was stirred for 2 days. The solvent was removed in vacuo andthe resulting residue was basified with 2N NaOH solution and the aqueouslayer was extracted with CH₂Cl₂ (2×30 mL). The combined organic layerswere dried over Na₂SO₄, filtered and the solvent was removed in vacuo toobtain (2S)-(4-amino-benzyl)-bicyclo[2.2.1]hept-2-yl-methyl-amineCompound 15f (0.54 g, 85% yield) as a thick yellow oil. MS m/e 231 (M⁺H,100%), which was used in the next step without further purification.

EDCI (0.33 mmol, 0.07 g) was added in one portion to a suspension ofCompound 15f (0.24 mmol, 0.06 g), 6-chloro-2H-chromene-3-carboxylic acidCompound 15g (0.22 mmol, 0.04 g) and HOBt (0.22 mmol, 0.03 g) in DMF(5.0 mL) at 0° C. The resulting suspension was warmed to r.t. and then acrystal of DMAP and Et₃N (0.65 mmol, 0.1 mL) was added and the reactionmixture was stirred overnight. The orange-yellow suspension was pouredin water and was extracted with EtOAc (25 mL). The organic layer waswashed with water (2×20 mL) followed by 5% NaOH solution (10 mL) andbrine. The organic layer was separated, dried over Na₂SO₄ and filtered.The solvent was removed in vacuo and the resulting residue was purifiedby preparative TLC (15:1 CH₂Cl₂/MeOH) to yield6-chloro-2H-chromene-3-carboxylic acid(2S)-{4-[(bicyclo[2.2.1]hept-2-yl-methyl-amino)-methyl]-phenyl}-amideCompound 15 h (0.06 g, 61%) as a pale yellow solid. MS m/e 423 (M⁺H,100%).

Iodomethane (0.5 mL) was added to a solution of Compound 15h (0.08 mmol,0.03 g) in CH₂Cl₂ (1.0 mL) at r.t. and the resulting solution wasallowed to stand overnight. A yellow precipitate was observed and thesolvent was removed in vacuo. The resulting yellow solid was washed withEt₂O to obtain Compound 110 (0.05 g, 96%) as a yellow solid. MS m/e 437(M⁺H, 100%).

Using the procedure of Example 15 and known appropriate reagents andstarting materials, other compounds of the present invention may beprepared including, (MS: Mass Spec data as MS m/e M⁺H):

Cpd Name MS 95{4-[(6-bromo-2H-chromene-3-carbonyl)-amino]-benzyl}-dimethyl- 471(tetrahydro-pyran-4-yl)-ammonium iodide 96{4-[(6-chloro-2H-chromene-3-carbonyl)-amino]-benzyl}-dimethyl- 427(tetrahydro-pyran-4-yl)-ammonium iodide 97{4-[(6-bromo-2H-chromene-3-carbonyl)-amino]-benzyl}-cyclohexyl- 469dimethyl-ammonium iodide 98{4-[(6-chloro-2H-chromene-3-carbonyl)-amino]-benzyl}-cyclohexyl- 425dimethyl-ammonium iodide 99(4-{[(6-bromo-2H-chromene-3-carbonyl)-amino]-methyl}-benzyl)-dimethyl-485 (tetrahydro-pyran-4-yl)-ammonium iodide 100{4-[(5,7-dichloro-2H-chromene-3-carbonyl)-amino]-benzyl}-dimethyl- 461(tetrahydro-pyran-4-yl)-ammonium iodide 101cyclohexyl-{4-[(5,7-dichloro-2H-chromene-3-carbonyl)-amino]-benzyl}- 459dimethyl-ammonium iodide 102{4-[(6,8-dichloro-2H-chromene-3-carbonyl)-amino]-benzyl}-dimethyl- 461(tetrahydro-pyran-4-yl)-ammonium iodide 103dimethyl-{4-[(6-methyl-2H-chromene-3-carbonyl)-amino]-benzyl}- 407(tetrahydro-pyran-4-yl)-ammonium iodide 104{4-[(6-methoxy-2H-chromene-3-carbonyl)-amino]-benzyl}-dimethyl- 423(tetrahydro-pyran-4-yl)-ammonium iodide 105cyclohexyl-dimethyl-{4-[(6-methyl-2H-chromene-3-carbonyl)-amino]- 405benzyl}-ammonium iodide 106cyclohexyl-{4-[(6-methoxy-2H-chromene-3-carbonyl)-amino]-benzyl}- 421dimethyl-ammonium iodide 107cyclohexyl-{4-[(6,8-dichloro-2H-chromene-3-carbonyl)-amino]-benzyl}- 459dimethyl-ammonium iodide 108(2R)-{4-[(6-chloro-2H-chromene-3-carbonyl)-amino]-benzyl}-dimethyl- 427(tetrahydro-furan-2-ylmethyl)-ammonium iodide 109(2S)-{4-[(6-chloro-2H-chromene-3-carbonyl)-amino]-benzyl}-dimethyl- 427(tetrahydro-furan-2-ylmethyl)-ammonium iodide 111bicyclo[2.2.1]hept-2-yl-{4-[(6,8-dichloro-2H-chromene-3-carbonyl)-amino]-471 benzyl}-dimethyl-ammonium iodide 112dimethyl-{4-[(8-methyl-2H-chromene-3-carbonyl)-amino]-benzyl}- 407(tetrahydro-pyran-4-yl)-ammonium iodide 113cyclohexyl-dimethyl-{4-[(8-methyl-2H-chromene-3-carbonyl)-amino]- 405benzyl}-ammonium iodide 114{4-[(6-chloro-8-methyl-2H-chromene-3-carbonyl)-amino]-benzyl}- 439cyclohexyl-dimethyl-ammonium iodide 115{4-[(6-chloro-8-methyl-2H-chromene-3-carbonyl)-amino]-benzyl}- 441dimethyl-(tetrahydro-pyran-4-yl)-ammonium iodide 116cyclohexyl-{4-[(7,8-dichloro-2H-chromene-3-carbonyl)-amino]-benzyl}- 459dimethyl-ammonium iodide 117bicyclo[2.2.1]hept-2-yl-{4-[(6-chloro-8-methyl-2H-chromene-3-carbonyl)-451 amino]-benzyl}-dimethyl-ammonium iodide 118{4-[(6-chloro-8-methyl-2H-chromene-3-carbonyl)-amino]-benzyl}- 453cycloheptyl-dimethyl-ammonium iodide 119{4-[(6-chloro-8-methyl-2H-chromene-3-carbonyl)-amino]-benzyl}- 425cyclopentyl-dimethyl-ammonium iodide 120{4-[(6-chloro-8-methyl-2H-chromene-3-carbonyl)-amino]-benzyl}- 443dimethyl-(tetrahydro-thiophen-3-yl)-ammonium iodide 121(4-{[(6-chloro-8-methyl-2H-chromene-3-carbonyl)-amino]-methyl}-benzyl)-455 dimethyl-(tetrahydro-pyran-4-yl)-ammonium iodide 122{4-[(6,8-dichloro-2H-chromene-3-carbonyl)-amino]-benzyl}-dimethyl- 463(tetrahydro-thiophen-3-yl)-ammonium iodide 123cyclohexyl-{4-[(6-fluoro-2H-chromene-3-carbonyl)-amino]-benzyl}- 409dimethyl-ammonium iodide 124cyclohexyl-{4-[(5-fluoro-2H-chromene-3-carbonyl)-amino]-benzyl}- 409dimethyl-ammonium iodide 125cyclohexyl-dimethyl-{4-[(6-trifluoromethyl-2H-chromene-3-carbonyl)- 459amino]-benzyl}-ammonium iodide 126cyclohexyl-{4-[(8-fluoro-2H-chromene-3-carbonyl)-amino]-benzyl}- 409dimethyl-ammonium iodide 127cyclohexyl-dimethyl-{4-[(7-methyl-2H-chromene-3-carbonyl)-amino]- 405benzyl}-ammonium iodide 128cyclohexyl-{4-[(7-methoxy-2H-chromene-3-carbonyl)-amino]-benzyl}- 421dimethyl-ammonium iodide 129{4-[(6-tert-butyl-2H-chromene-3-carbonyl)-amino]-benzyl}-cyclohexyl- 447dimethyl-ammonium iodide 130dimethyl-(tetrahydro-thiophen-3-yl)-{4-[(6-trifluoromethyl-2H-chromene-3-463 carbonyl)-amino]-benzyl}-ammonium iodide 131{4-[(5-fluoro-2H-chromene-3-carbonyl)-amino]-benzyl}-dimethyl- 413(tetrahydro-thiophen-3-yl)-ammonium iodide 132{4-[(6-fluoro-2H-chromene-3-carbonyl)-amino]-benzyl}-dimethyl- 413(tetrahydro-thiophen-3-yl)-ammonium iodide 133cyclohexyl-dimethyl-{4-[(5-trifluoromethyl-2H-chromene-3-carbonyl)- 459amino]-benzyl}-ammonium iodide 134cyclohexyl-dimethyl-{4-[(8-trifluoromethyl-2H-chromene-3-carbonyl)- 459amino]-benzyl}-ammonium iodide 135{4-[(3H-benzo[f]chromene-2-carbonyl)-amino]-benzyl}-dimethyl- 443(tetrahydro-pyran-4-yl)-ammonium iodide 1361-{4-[(3H-benzo[f]chromene-2-carbonyl)-amino]-benzyl}-1-methyl- 399pyrrolidinium iodide 137{4-[(3H-benzo[f]chromene-2-carbonyl)-amino]-benzyl}-cyclohexyl- 441dimethyl-ammonium iodide 138{4-[(3H-benzo[f]chromene-2-carbonyl)-amino]-benzyl}-dimethyl- 459(tetrahydro-thiopyran-4-yl)-ammonium iodide 1394-{4-[(3H-benzo[f]chromene-2-carbonyl)-amino]-benzyl}-4-methyl- 415morpholin-4-ium iodide 140{4-[(3H-benzo[f]chromene-2-carbonyl)-amino]-benzyl}-dimethyl- 457(tetrahydro-pyran-4-ylmethyl)-ammonium iodide 141(4-{[(3H-benzo[f]chromene-2-carbonyl)-amino]-methyl}-benzyl)-dimethyl-457 (tetrahydro-pyran-4-yl)-ammonium iodide

EXAMPLE 16{4-[(7,8-dichloro-2,3-dihydro-benzo[b]oxepine-4-carbonyl)-amino]-benzyl}-dimethyl-(tetrahydro-pyran-4-yl)-ammoniumiodide (Cpd 167)

Potassium carbonate (27.5 mmol, 3.75 g) and sodium iodide (0.3333 mmol,0.0500 g) were added to a reaction mixture of 3,4-dichloro-phenolCompound 16a (30.25 mmol, 4.93 g) and 4-bromo-butyric acid ethyl esterCompound 16b (27.5 mmol, 5.36 g) in acetone (60 mL). The reactionmixture was stirred overnight at room temperature. TLC analysis (4:1hexane:EtOAc) showed no formation of product. The reaction mixture wasrefluxed for 3 hrs and TLC analysis (4:1 hexane:EtOAc) showed trace ofstarting material Compound 16a. The reaction mixture was refluxedovernight, then basified with 1N NaOH solution and extracted withCH₂Cl₂. The organics were dried over MgSO₄. The drying agent wasfiltered and the solvent was removed in vacuo to yield4-(3,4-dichloro-phenoxy)-butyric acid ethyl ester Compound 16c (6.9 g,90.7%) as a pale pink oil, which was used in the next step withoutfurther purification. ¹H NMR (300 MHz, CDCl₃) δ 1.21-1.31 (q, 3H),2.12-2.25 (m, 2H), 2.45-2.53 (t, 2H), 3.95-4.02 (t, 2H), 4.10-4.20 (q,2H), 6.71-6.78 (dd, 1H), 6.94-6.96 (d, 1H), 7.28-7.31 (d, 1H).

A 1N NaOH solution (20 mL) was added to a solution of Compound 16c (7.22mmol, 2.00 g) in THF (20 mL) and MeOH (10 mL). The reaction mixturestirred overnight at room temperature. The THF and MeOH were removed invacuo and the remaining aqueous solution was acidified with 1N HCl. Aprecipitate was collected and dried in a vacuum oven overnight to yield4-(3,4-dichloro-phenoxy)-butyric acid Compound 16d (1.65 g, 92%) as awhite solid, which was used in the next step without furtherpurification. ¹H NMR (300 MHz, CDCl₃) δ 2.05-2.18 (m, 2H), 2.52-2.60 (t,2H), 3.95-4.05 (t, 2H), 6.70-6.79 (dd, 1H), 6.95-7.12 (d, 1H), 7.24-7.35(t, 1H).

Polyphosphoric acid (10 equivalents by weight, 51.2 g) was added to asolution of Compound 16d (0.0206 mol, 5.12 g) in toluene (51.5 mL). Themixture was heated to between 95 and 100° C. (bath temperature). Thereaction mixture was allowed to cool to room temperature and poured intoa beaker of ice water. The aqueous layer was extracted with Et₂O. Theorganics were washed with water and dried with MgSO₄. The drying agentwas filtered and the solvent was removed in vacuo; yielding a brownsolid (3.7 g), which was purified by flash column chromatography (2%EtOAc/Hexane to 10% EtoAc/Hexane) to yield7,8-dichloro-3,4-dihydro-2H-benzo[b]oxepin-5-one Compound 16e (1.52 g,32%) as a tan solid.

¹H NMR (300 MHz, CDCl₃) δ 2.18-2.28 (m, 2H), 2.84-2.95 (t, 2H),4.21-4.32 (t, 2H), 7.18 (s, 1H), 7.80 (s, 1H).

Sodium hydride (60% dispersion in mineral oil) (7.754 mmol, 0.3102 g)and dimethyl carbonate (37.41 mmol, 3.15 mL) were added to a reactionvessel. Compound 16e (3.877 mmol, 1.0 g) was dissolved in dimethylcarbonate (2 mL) and added dropwise to the vessel. The mixture wasrefluxed for 2 hrs. TLC analysis showed a trace of the starting materialCompound 16e and the reaction mixture was allowed to cool and stirredovernight at room temperature. 2N HCl solution (25 mL) was added to themixture, which was then extracted with EtOAc. The organics were driedover MgSO₄ and filtered. The solvent was removed in vacuo to yield abrown solid (1.27 g) which was taken up in Et₂O to yield a tanprecipitate. TLC analysis (30% Et₂O/Hexane) showed the precipitate to bepure (0.800 g). The remainder of the brown solid was purified by flashcolumn chromatography (30% Et₂O/Hexane) to yield7,8-dichloro-5-oxo-2,3,4,5-tetrahydro-benzo[b]oxepine-4-carboxylic acidmethyl ester Compound 16f (0.231 g) as a tan solid. ¹H NMR ofprecipitate (300 MHz, CDCl₃) δ 2.68-2.75 (t, 2H), 3.85 (s, 3H),4.32-4.38 (t, 2H), 7.12 (s, 1H), 8.08 (s, 1H), 13.15 (s, 1H).

A solution of Compound 16f (1.713 mmol, 0.4951 g) in CH₂Cl₂ (5 mL) wascooled to −15° C. while stirring. MeOH (0.75 mL) was added, followed bysodium borohydride (2.213 mmol, 0.0837 g) in two portions. The reactionmixture was stirred at −10° C. for 1 hr. TLC analysis (30% Et₂O/Hexane)of the reaction mixture showed complete formation of product, with notrace of starting material. The reaction mixture was washed with waterand dried over MgSO₄. The drying agent was filtered and the solvent wasremoved in vacuo yielding7,8-dichloro-5-hydroxy-2,3,4,5-tetrahydro-benzo[b]oxepine-4-carboxylicacid methyl ester Compound 16g (0.488 g, 97.8%) as a yellow oil, whichwas used in the next step without further purification. ¹H NMR (400 MHz,CDCl₃) δ 2.02-2.14 (m, 3H), 2.30-2.40 (m, 1H), 3.65 (s, 3H), 3.96-4.09(m, 2H), 4.20-4.28 (m, 1H), 7.35 (s, 1H), 7.56 (s, 1H).

A solution of Compound 16g (1.676 mmol, 0.4880 g) in CH₂Cl₂ (8 mL) wascooled to 0° C. while stirring. Triethylamine (5.018 mmol, 0.70 mL) wasadded, followed by the dropwise addition of methanesulfonyl chloride(2.506 mmol, 0.19 mL). The reaction mixture was allowed to warm to roomtemperature and stirred overnight. TLC analysis (30% Et₂O/Hexane) showedformation of product with no trace of starting material. The mixture wascooled to 0° C. and DBU (6.052 nmol, 0.90 mL) was added dropwise. Thereaction mixture was warmed to room temperature and stirred for 30 mins.An aliquot of the reaction mixture was washed with water, dried overMgSO₄, filtered and the solvent was removed in vacuo.

NMR analysis showed complete formation of product. The remainder of themixture was washed with water and dried over MgSO₄. The drying agent wasfiltered and the solvent was removed in vacuo to provide7,8-dichloro-2,3-dihydro-benzo[b]oxepine-4-carboxylic acid methyl esterCompound 16h (0.408 g, 86.1%) as a yellow oil. ¹H NMR (400 MHz, CDCl₃) δ2.95-3.00 (t, 2H), 3.82 (s, 3H), 4.22-4.28 (t, 2H), 7.10 (s, 1H), 7.40(s, 1H), 7.48 (s, 1H).

A solution of Compound 16h (1.494 mmol, 0.408 g), THF (20 mL), MeOH (10mL) and a solution of, 1N NaOH (20 mL) was stirred at room temperatureovernight. The solvent was removed in vacuo and the resulting aqueoussolution was acidified with concentrated HCl until an off-whiteprecipitate formed. The solid was filtered and dried in a vacuum ovenovernight to provide7,8-dichloro-2,3-dihydro-benzo[b]oxepine-4-carboxylic acid Compound 16i(0.373 g, 96.4%). ¹H NMR (300 MHz, CDCl₃) δ 2.90-3.02 (t, 2H), 4.25-4.31(t, 2H), 7.10 (s, 1H), 7.42 (s, 1H), 7.51 (s, 1H).

EDCI (0.3293 mmol, 0.0631 g) was added in one portion to a solution ofCompound 16i (0.2138 mmol, 0.0554 g),(4-amino-benzyl)-methyl-(tetrahydro-pyran-4-yl)-amine Compound 1d (T10)(0.2459 mmol, 0.0541 g) and HOBT (0.2138 mmol, 0.0289 g) in DMF (6 mL)at 0° C. The mixture was warmed to room temperature and a catalyticamount of DMAP and triethylamine (0.6414 mmol, 0.09 mL) were added. Thereaction mixture was stirred overnight at room temperature, then waterwas added and the mixture was extracted with EtOAc. The organics werewashed with water, 1 N NaOH solution and brine and dried over MgSO₄. Thedrying agent was filtered and the solvent was removed in vacuo to yielda yellow oil which was purified by TLC prep plate (9:1 EtoAc:MeOH) toyield 7,8-dichloro-2,3-dihydro-benzo[b]oxepine-4-carboxylic acid(4-{[methyl-(tetrahydro-pyran-4-yl)-amino]-methyl}-phenyl)-amideCompound 16j (0.040 g, 41%) as a yellow solid. MS m/e 461 (M⁺H, 90%),(M⁺Na, 100%); ¹H NMR (300 MHz, CDCl₃) δ 1.58-1.80 (m, 5H), 2.16 (s, 3H),2.58-2.69 (m, 1H), 3.00-3.08 (m, 2H), 3.30-3.41 (m, 2H), 3.52 (s, 2H),4.00-4.08 (m, 2H), 4.25-4.31 (m, 2H), 7.02 (s, 1H), 7.10 (s, 1H),7.28-7.38 (t, 3H), 7.50-7.55 (d, 2H).

Iodomethane (0.0161 mol, 1.0 mL) was added to a solution of Compound 16j(0.0433 mmol, 0.020 g) in acetonitrile (2 mL), acetone (2 drops) anddichloromethane (2 drops) at room temperature and the resulting solutionwas stirred overnight. The solvent was removed in vacuo and theresulting orange solid was washed with Et₂O and dried in a vacuum ovenfor 12 hrs to provide Compound 167 (0.0123 g, 78.5%). MS m/e 475 (M⁺H,100%); MS m/e 477 (M⁺H, 75%).

Using the procedure of Example 16 and known appropriate reagents andstarting materials, other compounds of the present invention may beprepared including, (MS: Mass Spec data as MS m/e M⁺H):

Cpd Name MS 142(4-{[(3-bromo-8,9-dihydro-7H-benzocycloheptene-6-carbonyl)-amino]- 497methyl}-benzyl)-dimethyl-(tetrahydro-pyran-4-yl)-ammonium iodide 143{4-[(3-bromo-8,9-dihydro-7H-benzocycloheptene-6-carbonyl)-amino]- 483benzyl}-dimethyl-(tetrahydro-pyran-4-yl)-ammonium iodide 144{4-[(3-bromo-8,9-dihydro-7H-benzocycloheptene-6-carbonyl)-amino]- 481benzyl}-cyclohexyl-dimethyl-ammonium iodide 1451-{4-[(8,9-dihydro-7H-benzocycloheptene-6-carbonyl)-amino]-benzyl}-1-361 methyl-pyrrolidinium iodide 146cyclohexyl-{4-[(8,9-dihydro-7H-benzocycloheptene-6-carbonyl)-amino]- 403benzyl}-dimethyl-ammonium iodide 147{4-[(8,9-dihydro-7H-benzocycloheptene-6-carbonyl)-amino]-benzyl}- 405dimethyl-(tetrahydro-pyran-4-yl)-ammonium iodide 148(4-{[(8,9-dihydro-7H-benzocycloheptene-6-carbonyl)-amino]-methyl}- 419benzyl)-dimethyl-(tetrahydro-pyran-4-yl)-ammonium iodide 168cyclohexyl-{4-[(7,8-dichloro-2,3-dihydro-benzo[b]oxepine-4-carbonyl)-473 amino]-benzyl}-dimethyl-ammonium iodide 169bicyclo[2.2.1]hept-2-yl-{4-[(7,8-dichloro-2,3-dihydro-benzo[b]oxepine-4-485 carbonyl)-amino]-benzyl}-dimethyl-ammonium iodide 170(4-{[(7,8-dichloro-2,3-dihydro-benzo[b]oxepine-4-carbonyl)-amino]- 489methyl}-benzyl)-dimethyl-(tetrahydro-pyran-4-yl)-ammonium iodide 171{4-[(7,8-dichloro-2,3-dihydro-benzo[b]oxepine-4-carbonyl)-amino]- 477benzyl}-dimethyl-(tetrahydro-thiophen-3-yl)-ammonium iodideBiological Activity

Compound of the invention were subjected to various representativebiological tests. The results of these tests are intended to illustratethe invention in a non-limiting fashion.

EXAMPLE 17 MCP-1 Receptor Binding Assay in THP-1 Cells

THP-1 cells were obtained from American Type Culture Collection(Manassas, Va., USA). The THP-1 cells were grown in RPMI-1640supplemented with 10% fetal bovine serum in a humidified 5% CO₂atmosphere at 37° C. The cell density was maintained between 0.5×10⁶cells/mL.

THP-1 cells were incubated with 0.5 nM ¹²⁵I labeled MCP-1 (Perkin-ElmerLife Sciences, Inc. Boston, Mass.) in the presence of varyingconcentrations of either unlabeled MCP-1 (R&D Systems, Minneapolis,Minn.) or test compound for 2 hours at 30° C. in a 96 well plate. Cellswere then harvested onto a filter plate, dried, and 20 μL of Microscint20 was added to each well. Plates were counted in a TopCount NXT,Microplate Scintillation & Luminescence Counter (Perkin-Elmer LifeSciences, Inc. Boston, Mass.). Blank values (buffer only) weresubtracted from all values and drug treated values were compared tovehicle treated values. 1 μM cold MCP-1 was used for nonspecificbinding.

Table 1 lists IC₅₀ values for inhibition of MCP-1 binding to CCR2obtained for test compounds of the invention.

Table 2 lists inhibition values obtained for test compounds for MCP-1binding to CCR2. The inhibition values (%) were obtained at a testconcentration of 25 μM, unless indicated otherwise.

TABLE 1 Mean Ligand Binding (IC₅₀ μM) Cpd IC₅₀ (μM) 1 2.3 2 5.5 3 4.3 43.1 5 8.3 6 5 7 6.3 10 6.6 11 3.1 14 0.06 15 2 16 0.85 17 0.005; 0.01;0.009 18 1.8 19 0.005 20 0.16 21 0.15 22 1.6 23 0.9 24 0.29; 0.19 260.28 27 0.75 28 0.46 29 0.33 30 4.2 31 11.4 32 3.4 33 0.4 34 2.4 35 0.437 10.2 38 5 39 6.4 40 1.6 42 5.7 43 0.34 44 14.6 45 0.26 46 1.3 47 3.748 8.4 49 11.9 50 2.6 51 2.7 52 2.6 53 3.2 55 5.4 56 11.8 57 13.3 59 1061 2.7 63 3.5 66 1.7 67 2.5 68 1.5 69 6.4 70 0.11 71 0.17; 0.32 72 0.5873 0.44 74 1.5 75 0.02 76 0.16 78 13.4 79 0.31 80 0.03 81 3 82 0.05 831.2 84 3.2 85 5.2 86 6 87 3.8 88 0.87 90 0.22 91 4.1 93 0.9 94 1.3 950.18 96 0.11 97 0.45 98 0.27 100 0.52 101 1.1 102 0.19 103 0.82 104 1.9105 0.32 106 1.1 107 0.17 108 3.1 109 5.3 110 0.32 111 2 112 0.15 1130.53 114 0.06 115 0.05 116 0.6 117 0.12 118 0.2 119 0.1 120 0.03 1220.58 126 0.91 127 0.08 128 0.6 129 0.15 130 0.3 131 0.33 132 0.13 1333.3 134 0.47 135 0.91 136 4.6 137 0.08 138 2.4 139 9.8 140 1 143 0.007144 0.33 146 0.71 147 1.2 151 0.26 152 1.6 153 2.1 154 4.6 155 0.49 1560.18 157 0.69 158 0.4 159 0.19 160 0.29 162 1.6 163 0.62 164 0.2 165 3.3166 0.11 167 0.24 168 1 169 0.24 171 0.56 173 8.1 175 1.5 176 0.26 1770.17 178 8.8

TABLE 2 % Inhibition Ligand Binding Cpd % Inhibition 8 34 9 65 12 38 1329 25 15 36 49 41 87 58 53 60 20 62 9 64 63 65 61 77 53 89 29 92 54 9934 121 37 123 100 124 100 125 100 141 −21 142 −18 145 36 148 7 149 58150 27 161 33 170 36 172 17 174 52

EXAMPLE 18 MCP-1 Induced Chemotaxis in THP-1 Cells

MCP-1 induced chemotaxis was run in a 24-well chemotaxis chamber. MCP-1(0.01 μg/mL) was added to the lower chamber and 100 μL of THP-1 cells(1×10⁷ cell/mL) was added to the top chamber. Varying concentrations oftest compound were added to the top and bottom chambers. Cells wereallowed to chemotax for 3 hours at 37° C. and 5% CO₂. An aliquot of thecells which had migrated to the bottom chamber was taken and countedthen compared to vehicle.

Test compounds of the invention inhibited MCP-1 induced chemotaxis withIC₅₀ values of from about 10 μM to about 1 nM.

EXAMPLE 19 MCP-1 Induced Calcium Mobilization in THP-1 Cells

THP-1 cells were plated at a density of 8×10⁵ cells/ml (100 μL/well)into poly-D lysine coated clear bottom, black 96 well plates. The cellswere loaded with 5 μM fluo-3 for 45 minutes. The fluo-3 was washed offand cells were incubated with varying concentrations of test compoundfor 15 minutes. The change in [Ca²⁺]_(i) upon addition of 0.2 μM MCP-1is determined using FLIPR and compared to vehicle.

Test compounds of the invention inhibited MCP-1 induced influx of Ca²⁺ions with IC₅₀ values of from about 10 μM to about 1 nM.

EXAMPLE 20 Inhibition of Uveitis in Mice

The lipopolysaccharide (LPS bacterial endotoxin) induced uveitis mousemodel is used to test a compound of the invention for inhibition ofMCP-1 induced inflammation in the anterior of the eye (Tuaillon N, Shende F, Berger R B, Lu B, Rollins B J and Chan CC, MCP-1 expression inendotoxin-induced uveitis, Invest. Ophthalmol. Vis. Sci., 2002 May,43(5): 1493-8).

After intraocular injection directly into the anterior chamber of theeye with LPS, a measurable amount of MCP-1 is found in the eye's aqueoushumor within a few hours. The degree of inflammation is quantified bycounting the number of leukocytes within the aqueous humor of theanterior chamber (including a differential count), determining theprotein concentration in the aqueous humor and confirming the inhibitionof inflammation by histological examination.

Procedure

A test compound was dissolved in saline (5 mg/mL); and 10 μL (50 μg) wasapplied topically to the injected eye at 0, 4, and 8 hr relative to theLPS injection. The control group was treated topically with a salinevehicle (no test compound). One hour after the last dose of the compound(i.e., 9 hours post-injection), the mice were sacrificed and leukocyte,neutrophil and mononuclear cell counts and protein concentration insidethe eye were measured.

Results

In two trials, the compound inhibited leukocyte infiltration by 66%(±1%). The accumulation of protein was inhibited by 52% (±14%). Celldifferential counts indicated that neutrophil influx into the eye wasinhibited by 67% while mononuclear cell influx was inhibited by 40%.Histological examination confirmed the inhibition of cellular influx.

Based on the binding data for inhibition of MCP-1 induced inflammationand the data for inhibition of MCP-1 induced anterior uveitis, aneffective dose per day for a compound of the invention for treatinganterior uveitis is in a range of from about 50 μg to about 0.5 ng. Anembodiment of an effective dose for a compound of the invention for thetreatment of anterior uveitis is from about 5 μg to about 0.5 ng.Another embodiment of an effective dose for such treatment is from about1 μg to about 1 ng. Another embodiment of an effective dose is fromabout 0.5 μg to about 1 ng. An embodiment of an effective dose is alsofrom about 0.1 μg to about 1 ng.

EXAMPLE 21 Inhibition of Ovalbumin (Ova)-Induced Asthma in Mice

Test compounds of the present invention were active in two differentmodels of ovalbumin (OVA)-induced asthma in mice.

Mast Cell-Dependent Model

Mice were sensitized by i.p. injection with OVA in saline (10 μg) onalternate days (Day 0, 2, 4, 6, 8, 10, 12). Groups of mice were eachchallenged by intranasal injection of OVA (Day 40, 43, 46). Compound 17was administered by i.p. injection (30 mg/kg) on consecutive days (Day42, 43, 44, 45, 46). Compared to vehicle, leukocyte influx was inhibitedby 95% and 55% (in two separate assays), LTC₄ influx was inhibited by90% and IL-4 influx was inhibited by 85%.

Mast Cell-Independent Model

Mice were sensitized by i.p. injection of OVA emulsified in adjuvant(Day 1 and 14). Groups of mice were each challenged by intranasalinjection of OVA (Day 25, 26, 27). Compound 17 was administered by i.p.injection (10 and 30 mg/kg) before each intranasal challenge (Day 25,26, 27). Compared to vehicle, leukocyte influx was dose-dependentlyinhibited by 40% and 70%, respectively.

EXAMPLE 22 Inhibition of Ovalbumin-induced Allergic Rhinitis in Mice

BALB/c mice were sensitized by i.p. injection of OVA emulsified in alum(Day 0, 5, 14, 21). Groups of mice were each challenged by intranasalinjection of OVA (Day 22-35, 38).

Control group mice received an equal volume of vehicle by intranasalinjection. Nasal symptoms (number of sneezes and episodes of noserubbing by the front paws) were counted during the 5 min periodfollowing the last intranasal injection (Day 38).

Prophylactic Effect

Compound 17 (in PBS) was administered by intranasal injection (10 and 30μg/nostril) to both nostrils twice daily 1 hr and 6 hrs prior tointranasal challenge (Days 22-35), once per day prior to intranasalchallenge (Days 36, 37) then 1 hr and 6 hrs prior to intranasalchallenge (Day 38). The histamine receptor antagonist Astelin® was usedas a positive control.

Compared to vehicle, Compound 17 dose-dependently inhibited nasalsymptoms by 64/57% (sneezing/rubbing) and 82/71% (sneezing/rubbing),respectively. Compared to vehicle, the positive control inhibited nasalsymptoms by 51/89% (sneezing/rubbing).

Therapeutic Effect

The dosing of Compound 17 was delayed until the symptoms of rhinitis hadappeared (Day 29). Compound 17 (in PBS) was then administered byintranasal injection (10 μg/nostril) to both nostrils four times per dayprior to intranasal challenge (Days 29-38). The anti-histaminePyralimine and the mast cell-stabilizing agent Ketotifen were used aspositive controls.

Compared to vehicle, Compound 17 inhibited nasal symptoms by 0/42%(sneezing/rubbing). Compared to vehicle, Pyralimine and Ketotifeninhibited nasal symptoms by 60/85% (sneezing/rubbing) and 50/81%(sneezing/rubbing), respectively.

While the foregoing specification teaches the principles of the presentinvention, with examples provided for the purpose of illustration, itwill be understood that the practice of the invention encompasses all ofthe usual variations, adaptations and/or modifications as come withinthe scope of the following claims and their equivalents.

1. A compound which is a quaternary ammonium salt, the cation havingFormula (I)

or pharmaceutically acceptable forms of said compound thereof, wherein Ais carbonyl, thiocarbonyl or sulfonyl; X is a bond —or —CH═CH—; R₁ isselected from (1). aryl optionally substituted by one or more loweralkyl, —(CH₂)_(n)—CF₃, lower alkoxy, alkoxycarbonyl, cyano, halogen orphenyl optionally substituted by lower alkyl, —(CH₂)_(n)—CF3, loweralkoxy, alkoxycarbonyl, cyano or halogen; or (2). C₅-C₁₅ cycloalkyloptionally substituted by one or more lower alkyl, —(CH₂)_(n)—CF₃, loweralkoxy, aryl, halogen-substituted aryl, alkoxycarbonyl, cyano orhalogen; n is 0, 1, 2, 3, or 4; Y is a bond or —CH₂—; X₂ is —(CH₂)_(m)—wherein m is 1 or 2; R₂ is —N⁺(R4R₅)—ZR₃; Z is —(CH₂)_(p)— wherein p is0, 1 or 2; R₃ is selected from (1). aryl optionally substituted by oneor more lower alkyl, —(CH₂)_(n)—CF₃, lower alkoxy, aryl,halogen-substituted aryl, alkoxycarbonyl, cyano or halogen; (2). C₅-C₁₅cycloalkyl optionally substituted with one or more lower alkyl,—(CH₂)_(n)—CF₃, lower alkoxy, aryl, halogen-substituted aryl,alkoxycarbonyl, cyano or halogen; or (3). heterocyclyl optionallysubstituted with one or more lower alkyl, —(CH₂)_(n)—CF₃, lower alkoxy,aryl, halogen-substituted aryl, alkoxycarbonyl, cyano or halogen;wherein, when heterocyclyl is attached via a carbon atom ring member anda heteroatom ring member is adjacent to said carbon atom, then p is 1 or2; R₄ and R₅ are each individually lower alkyl or lower alkenyl; oralternatively, R₄ and R₅ combine with the nitrogen atom of Formula (I)to form a heterocyclyl ring of 5 to 9 total ring atoms optionallycontaining one of an oxygen or sulfur ring atom, wherein theheterocyclyl ring nitrogen atom is substituted with one of lower alkylor lower alkenyl to form a quaternary salt, and wherein —ZR₃ is absentand the heterocyclyl ring is optionally substituted with aryl optionallysubstituted with one or more lower alkyl, —(CH₂)_(n)—CF₃, lower alkoxy,aryl, halogen-substituted aryl, alkoxycarbonyl, cyano or halogen, saidheterocyclyl for ring R₃ being tetrahydropyran, tetrahydrofuran,tetrahydrothiophene, tetrahydrothiopyran, pyrrolidine, morpholine,thiomorpholine, piperidine, thiophene or piperazine.
 2. The compound ofclaim 1, wherein A is carbonyl; X is a bond; R₁ is selected from arylsubstituted by one or more lower alkyl or halogen, and C₅-C₁₅ cycloalkyloptionally substituted by one or more halogen, Y is a bond; X₂ is —CH₂—;R₂ is —N⁺(R₄R₅)—R₃; R₃ is selected from C₅-C₁₅ cycloalkyl orheterocyclyl and R₄ and R₅ are each individually lower alkyl.
 3. Thecompound of claim 1, wherein A is carbonyl, X is a bond, R₁ is aryloptionally substituted by one or more halogen, Y is a bond, X₂ is —CH₂—,R₂ is —N⁺(R₄R₅)—R₃, R₃ is heterocyclyl and R₄ and R₅ are eachindividually lower alkyl.
 4. The compound of claim 1, wherein A iscarbonyl.
 5. The compound of claim 1, wherein R₁ is selected from (1).aryl optionally substituted by one or more lower alkyl, —(CH₂)_(n)—CF₃,lower alkoxy, cyano, halogen or phenyl optionally substituted by loweralkyl, —(CH₂)_(n)——CF₃, lower alkoxy, cyano or halogen; and (2). C₅-C₁₅cycloalkyl optionally substituted by one or more lower alkyl,—(CH₂)_(n)—CF₃, lower alkoxy, cyano or halogen.
 6. The compound of claim1, wherein n is
 0. 7. The compound of claim 1, wherein p is 0 or
 1. 8.The compound of claim 1, wherein R₃ is C₅-C₁₅ cycloalkyl orheterocyclyl; wherein, when heterocyclyl is attached via a carbon atomring member and a heteroatom ring member is adjacent to said carbonatom, then p is
 1. 9. The compound of claim 1, wherein R₄ and R₅ areeach individually lower alkyl or lower allyl.
 10. The compound of claim1, wherein R₄ and R₅ combine with the nitrogen atom of Formula (I) toform a heterocyclyl ring of 5 to 9 total ring atoms optionallycontaining one of an oxygen or sulfur ring atom, wherein theheterocyclyl ring nitrogen atom is substituted with lower alkyl to forma quaternary salt, and wherein —ZR₃ is absent and the heterocyclyl ringis optionally substituted with one or more lower alkyl, —(CH₂)_(n)—CF₃,lower alkoxy, cyano or halogen.
 11. The compound of claim 1, wherein R₄and R₅ combine with the nitrogen atom of Formula (I) to form aheterocyclyl ring of 5 to 9 total ring atoms optionally containing oneof an oxygen or sulfur ring atom, wherein the heterocyclyl ring nitrogenatom is substituted with lower alkyl to form a quaternary salt, andwherein —ZR₃ is absent and the heterocyclyl ring is optionallysubstituted with aryl optionally substituted with lower alkoxy.
 12. Acompound which is a quaternary ammonium salt, or pharmaceuticallyacceptable forms thereof, the cation of said compound selected from


13. A composition comprising an effective amount of the compound ofclaim 1 and a pharmaceutically acceptable carrier.
 14. The compositionof claim 13 selected from a topically applied composition, anintranasally applied composition or an ocularly applied composition. 15.A process for preparing the composition of claim 13 comprising the stepof admixing said compound with a pharmaceutically acceptable carrier.16. A method for treating CCR2 mediated inflammation in a subject inneed thereof comprising administering to the subject an effective amountof the compound of claim 1 or pharmaceutically acceptable form of saidcompound.
 17. The method of claim 16 wherein the effective amount isfrom about 0.001 mg/kg/day to about 300 mg/kg/day.
 18. The method ofclaim 16, wherein the CCR2 mediated inflammation is associated withelevated MCP-1 expression or MCP-1 over expression, or is aninflammatory condition that accompanies syndromes, disorders or diseasesassociated with elevated MCP-1 expression or MCP-1 overexpression. 19.The method of claim 16, wherein the CCR2 mediated inflammation isassociated with ophthalmic disorders, atherosclerosis, rheumatoidarthritis, psoriasis, psoriatic arthritis, atopic dermatitis, multiplesclerosis, Crohn's Disease, ulcerative colitis, nephritis, organallograft rejection, fibroid lung, renal insufficiency, diabetes anddiabetic complications, diabetic nephropathy, diabetic retinopathy,diabetic retinitis, diabetic microangiopathy, tuberculosis, chronicobstructive pulmonary disease, sarcoidosis, invasive staphyloccocia,inflammation after cataract surgery, allergic rhinitis, allergicconjunctivitis, chronic utricaria, asthma, periodontal diseases,diastolic cardiomyopathies, cardiac infarction, myocarditis, chronicheart failure, angiostenosis, restenosis, reperfusion disorders,glomerulonephritis, solid tumors and cancers, chronic lymphocyticleukemia, chronic myelocytic leukemia, multiple myeloma, malignantmyeloma or Hodgkin's disease.
 20. The method of claim 16, wherein themethod comprises treating CCR2 mediated inflammation associated withophthalmic disorders, rheumatoid arthritis, psoriasis, psoriaticarthritis, atopic dermatitis, chronic obstructive pulmonary disease,allergic rhinitis, asthma, or periodontal diseases in a subject in needthereof by administering to the subject an effective amount of saidcompound or pharmaceutically acceptable form of said compound.
 21. Themethod of claim 20, wherein the ophthalmic disorder is selected fromuveitis or allergic conjunctivitis and the periodontal disease isselected from periodontitis and gingivitis.
 22. The method of claim 21,wherein uveitis is acute, recurring or chronic uveitis.
 23. The methodof claim 21, wherein uveitis is anterior uveitis, intermediate uveitis,posterior uveitis or panuveitis.
 24. The method of claim 16, wherein themethod comprises treating CCR2 mediated inflammation associated withacute uveitis, recurring uveitis, chronic uveitis, allergicconjunctivitis, rheumatoid arthritis, psoriasis, psoriatic arthritis,atopic dermatitis, chronic obstructive pulmonary disease, allergicrhinitis, asthma, periodontitis or gingivitis in a subject in needthereof by administering to the subject an effective amount of saidcompound or pharmaceutically acceptable form of said compound.
 25. Themethod of claim 16, wherein the method comprises treating a CCR2mediated inflammation in a subject in need thereof by administering tothe subject an effective amount of said compound or pharmaceuticallyacceptable form of said compound in a combination therapy with one ormore anti-inflammatory agents, anti-infective agents orimmunosuppressive agents.
 26. The compound according to claim 1 whereinthe cation of Formula (I) is:


27. The method of claim 16 wherein the CCR2 mediated inflammation isassociated with rhinitis, uveitis, asthma, periodontitis, or gingivitis.28. The method according to claim 16 wherein the cation of Formula (I)is


29. The method according to claim 16 wherein the CCR2 mediatedinflammation is associated with periodontitis or gingivitis.
 30. Themethod of claim 19 wherein the ophthalmic disorder is uveitis, theasthma is allergic asthma, the periodontal disease is periodontitis orgingivitis and the solid tumor and cancers are carcinomas of thebladder, breast, cervix, colon, lung, prostate or stomach.
 31. Themethod of claim 20 wherein the asthma is allergic asthma.
 32. The methodof claim 24 wherein the asthma is allergic asthma.
 33. The method ofclaim 16 wherein the CCR2 mediated inflammation is associated withophthalmic disorder, asthma, periodontal disease, or allergic rhinitis.34. The method according to claim 33 wherein the periodontal disease isgingivitis or periodontitis and the ophthalmic disorder is uveitis. 35.The method according to claim 27 wherein rhinitis is allergic rhinitis.